U.S. patent application number 10/513552 was filed with the patent office on 2006-09-14 for 1,1-and 1,2-bisphosphonates as apoliprotein e modulators.
Invention is credited to Craig Leigh Bentzen, Emanuele Burattini, Vinh Van Diep, Yves Guyon-Gellin, Imber Flores Montes, Eric Joseph Neisor, Lan Mong Nguyen, Anne Perez, Hieu Trung Pham, Carlo Severi, Jean-Luc Thuillard.
Application Number | 20060205692 10/513552 |
Document ID | / |
Family ID | 29420566 |
Filed Date | 2006-09-14 |
United States Patent
Application |
20060205692 |
Kind Code |
A1 |
Montes; Imber Flores ; et
al. |
September 14, 2006 |
1,1-and 1,2-Bisphosphonates as apoliprotein e modulators
Abstract
The present invention relates to methods of use 1,1- and
1,2-bisphosphonate compounds to modulate apolipoprotein E levels
and use of such compounds in therapy, including cardiovascular and
neurological disease states.
Inventors: |
Montes; Imber Flores; (Le
Lignon, CH) ; Pham; Hieu Trung; (Tannay, CH) ;
Nguyen; Lan Mong; (Nyon, CH) ; Diep; Vinh Van;
(Vetraz-Monthoux, FR) ; Burattini; Emanuele;
(Geveva, CH) ; Severi; Carlo; (Le Lignon, CH)
; Neisor; Eric Joseph; (Nyon, CH) ; Perez;
Anne; (Versoix, CH) ; Thuillard; Jean-Luc;
(Saint Cergue, CH) ; Guyon-Gellin; Yves;
(Annemasse, FR) ; Bentzen; Craig Leigh;
(Bogis-Bossey, CH) |
Correspondence
Address: |
MCDONNELL BOEHNEN HULBERT & BERGHOFF LLP
300 S. WACKER DRIVE
32ND FLOOR
CHICAGO
IL
60606
US
|
Family ID: |
29420566 |
Appl. No.: |
10/513552 |
Filed: |
May 9, 2003 |
PCT Filed: |
May 9, 2003 |
PCT NO: |
PCT/US03/14727 |
371 Date: |
October 21, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60379835 |
May 11, 2002 |
|
|
|
Current U.S.
Class: |
514/80 ; 514/102;
514/89; 546/22; 548/415; 558/158 |
Current CPC
Class: |
C07F 9/404 20130101;
C07F 9/58 20130101; C07F 9/6533 20130101; C07F 9/59 20130101; C07F
9/5728 20130101; C07F 9/4025 20130101; C07F 9/572 20130101; C07F
9/5765 20130101 |
Class at
Publication: |
514/080 ;
514/089; 514/102; 546/022; 548/415; 558/158 |
International
Class: |
A61K 31/675 20060101
A61K031/675; A61K 31/663 20060101 A61K031/663; C07F 9/58 20060101
C07F009/58; C07F 9/572 20060101 C07F009/572; C07F 9/28 20060101
C07F009/28 |
Claims
1. A bisphosphonate derivative of the formula: ##STR40## wherein A
is hydroxy, aryl, heterocycle or --NR.sup.3R.sup.4 wherein R.sup.3
and R.sup.4 are independently hydrogen or C.sub.1-C.sub.4 alkyl; L
is --(CH.sub.2)--, --(CH.sub.2).sub.pO(CH.sub.2).sub.q--,
--(CH.sub.2).sub.pNR.sup.5(CH.sub.2).sub.q-- or
--(CH.sub.2).sub.pNHCO(CH.sub.2).sub.q--, wherein R.sup.5 is
hydrogen, C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.3 cyanoalkyl; and
m, p and q are an integer from 0 to 6; R.sup.1 and R.sup.2 are
independently hydrogen or C.sup.1-C.sub.6 alkyl; is a single or a
double bond; M is (CH.sub.2).sub.n or (CH.dbd.CH).sub.u--CH.dbd.
where n is an integer from 0 to 3 and u is 0 or 1; B is H or
C.sub.1-C.sub.4 alkyl group and w is 0 when is a double bond and w
is 1 when is a single bond or when M is (CH.sub.2).sub.n and n is
0; s is 0 or 1; Z.sup.1and Z.sup.2 are independently hydrogen,
C.sub.1-C.sub.4 alkyl, or C.sub.1-C.sub.4 alkoxy; with the proviso
that if n is 1, 2 or 3 or M is (CH.dbd.CH).sub.u--CH=, then s is 1
and/or A-L-O, Z.sup.1 and Z.sup.2 are not all independently H,
alkyl or alkoxy; or a pharmaceutically acceptable salt thereof.
2. The bisphosphonate derivative of claim 1, wherein and Z.sup.1
and Z.sup.2 are hydrogen and n is 0 or 1 or u is 0.
3. The bisphosphonate derivative of claim 1, wherein L is M is
(CH.sub.2).sub.n and n is 2 or 3.
4. The bisphosphonate derivative of claim 3, wherein A is
pyridin-2-yl, pyridin-3-yl, pyrrolidino, succinomido, piperidino,
morpholino, phthalimido, phenyl, p-cyanophenyl or
N,N'-(2-cyanoethyl)phenyl-amino.
5. The bisphosphonate derivative of claim 4, wherein A is
pyridin-2-yl, pyridin-3-yl, phthalimido, or
N,N'-(2-cyanoethyl)phenyl-amino.
6. The bisphosphonate derivative of claim 1 or claim 2, wherein
R.sup.1 and R.sup.2 the same and are methyl, ethyl or
isopropyl.
7. The bisphosphonate derivative of claim 1, wherein said
bisphosphonate derivative is tetraethyl
1-[4-(3-N-phthalimido-propoxy)-phenyl]-methylidene-1,1-bisphosphonate
8. The bisphosphonate derivative of claim 1, wherein said
bisphosphonate derivative is tetraethyl
1-[4-(3-N-phthalimido-propoxy)-phenyl]-ethylidene-1,2-bisphosphonate
9. The bisphosphonate derivative of claim 1, wherein said
bisphosphonate derivative is tetraethyl
1-{4-[3-(methyl-pyridin-2-yl-amino)-propoxy]-phenyl}-methylidene-1,1-bisp-
hosphonate.
10. The bisphosphonate derivative of claim 1, wherein said
bisphosphonate derivative is tetraethyl
1-(4-{2-[(2-cyano-ethyl)-phenyl-amino]-ethoxy}-phenyl)-methylidene-1,1-bi-
sphosphonate.
11. A method of modulating the production of apoE by an apoE
producing cell, comprising contacting said apoE producing cell with
an effective amount of a bisphosphonate of the formula: ##STR41##
wherein Y is hydrogen, hydroxy, halo, aryl, aryloxy,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy or A-LO; A is
hydroxy, aryl, heterocycle or --NR.sup.3R.sup.4 wherein R.sup.3 and
R.sup.4 are independently hydrogen or C.sub.1-C.sub.4 alkyl; L is
--(CH.sub.2).sub.m--, --(CH.sub.2).sub.pO(CH.sub.2).sub.q--,
--(CH.sub.2).sub.pNR.sup.5(CH.sub.2).sub.q-- or
--(CH.sub.2).sub.pNHCO(CH.sub.2).sub.q--, wherein R.sup.5 is
hydrogen, C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.3 cyanoalkyl; and
m, p and q are an integer from 0 to 6; R.sup.1 and R.sup.2 are
independently hydrogen or C.sub.1-C.sub.6 alkyl; is a single or a
double bond; M is (CH.sub.2).sub.n or (CH.dbd.CH).sub.u--CH.dbd.
where n is an integer from 0 to 3 and u is 0 or 1; B is H or
C.sub.1-C.sub.4 alkyl group and w is 0 when is a double bond and w
is 1 when is a single bond or when M is (CH.sub.2).sub.n and n is
0; s is 0 or 1; Z.sup.1 and Z.sup.2 are independently hydrogen,
C.sub.1-C.sub.4 alkyl, or C.sub.1-C.sub.4 alkoxy; with the proviso
that if n is 1, 2 or 3 or M is (CH.dbd.CH).sub.u--CH.dbd., then s
is 1 and/or Y, Z.sup.1 and Z.sup.2 are not all independently H,
hydroxy, alkyl or alkoxy; or a pharmaceutically acceptable salt
thereof.
12. The bisphosphonate derivative of claim 11, wherein Z.sup.1 and
Z.sup.2 are hydrogen and n is 0 or 1 or u is 0.
13. The bisphosphonate derivative of claim 11, wherein Y is
A-L-O.
14. The bisphosphonate derivative of claim 13, wherein A is
pyridin-2-yl, pyridin-3-yl, pyrrolidino, succinomido, piperidino,
morpholino, phthalimido, phenyl, p-cyanophenyl or
N,N'-(2-cyanoethyl)phenyl-amino.
15. The bisphosphonate derivative of claim 14, wherein A is
pyridin-2-yl, pyridin-3-yl, phthalimido, or
N,N'-(2-cyanoethyl)phenyl-amino.
16. The method of claim 11 or claim 2, wherein R.sup.1 and R.sup.2
the same and are methyl, ethyl or isopropyl.
17. The method of claim 11, wherein the bisphosphonate derivative
is tetraethyl
1-[4-(3-N-phthalimido-propoxy)-phenyl]-methylidene-1,1-bisphosphonate,
tetraethyl
1-[4-(3-N-phthalimido-propoxy)-phenyl]-ethylidene-1,2-bisphosphonate,
tetraethyl
1-{4-[3-(methyl-pyridin-2-yl-amino)-propoxy]-phenyl}-methylidene-1,1-bisp-
hosphonate or tetraethyl
1-(4-{2-[(2-cyano-ethyl)-phenyl-amino]-ethoxy}-phenyl)-methylidene-1,1-bi-
sphosphonate.
18. The method of claim 11, wherein said modulating the production
of apoE increases the production of apoE.
19. The method of claim 11, wherein said modulating the production
of apoE decreases the production of apoE.
20. A method of modulating apoE levels in a patient in need of such
treatment, comprising administration of an effective amount of a
bisphosphonate derivative of the formula: ##STR42## wherein Y is
hydrogen, hydroxy, halo, aryl, aryloxy, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxy or A-L-O; A is hydroxy, aryl, heterocycle or
--NR.sup.3R.sup.4 wherein R.sup.3 and R.sup.4 are independently
hydrogen or C.sub.1-C.sub.4 alkyl; L is --(CH.sub.2).sub.m--,
--(CH.sub.2).sub.pO(CH.sub.2).sub.q--,
--(CH.sub.2).sub.pNR.sup.5(CH.sub.2).sub.q-- or
--(CH.sub.2).sub.pNHCO(CH.sub.2).sub.q--, wherein R.sup.5 is
hydrogen, C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.3 cyanoalkyl; and
m, p and q are an integer from 0 to 6; R.sup.1 and R.sup.2 are
independently hydrogen or C.sub.1-C.sub.6 alkyl; is a single or a
double bond; M is (CH.sub.2).sub.n or (CH.dbd.CH).sub.u--CH.dbd.
where n is an integer from 0 to 3 and u is 0 or 1; B is H or
C.sub.1-C.sub.4 alkyl group and w is 0 when is a double bond and w
is 1 when is a single bond or when M is (CH.sub.2).sub.n and n is
0; s is 0 or 1; Z.sup.1 and Z.sup.2 are independently hydrogen,
C.sub.1-C.sub.4 alkyl, or C.sub.1-C.sub.4 alkoxy; with the proviso
that if n is 1, 2 or 3 or M is (CH.dbd.CH).sub.u--CH=, then s is 1
and/or Y, Z.sup.1 and Z.sup.2 are not all independently H, hydroxy,
alkyl or alkoxy; or a pharmaceutically acceptable salt thereof
21. The bisphosphonate derivative of claim 20, Z.sub.1 and Z.sup.2
are hydrogen and n is 0 or 1 or u is 0.
22. The bisphosphonate derivative of claim 21, wherein Y is
A-L-O.
23. The bisphosphonate derivative of claim 22, wherein A is
pyridin-2-yl, pyridin-3-yl, pyrrolidino, succinomido, piperidino,
morpholino, phthalimido, phenyl, p-cyanophenyl or
N,N'-(2-cyanoethyl)phenyl-amino.
24. The bisphosphonate derivative of claim 23, wherein A is
pyridin-2-yl, pyridin-3-yl, phthalimido, or
N,N'-(2-cyanoethyl)phenyl-amino.
25. The method of claim 20 or claim 2, wherein R.sup.1 and R.sup.2
the same and are methyl, ethyl or isopropyl.
26. The method of claim 20, wherein the bisphosphonate derivative
is tetraethyl
1-[4(3-N-phthalimido-propoxy)-phenyl]-methylidene-1,1-bisphosphonate,
tetraethyl
1-[4-(3-N-phthalimido-propoxy)-phenyl]-ethylidene-1,2-bisphosphonate,
tetraethyl
1-{4-[3-(methyl-pyridin-2-yl-amino)-propoxy]-phenyl}-methylidene-1,1-bisp-
hosphonate or tetraethyl
1-(4-{2-[(2-cyano-ethyl)-phenyl-amino]-ethoxy}-phenyl)-methylidene-1,1-bi-
sphosphonate.
27. The method of claim 20, wherein said modulation of said apoE
levels in said patient comprises increasing said apoE levels.
28. The method of claim 27, wherein said patient is suffering from
atherosclerosis, Alzheimer's disease, macular degeneration,
retinitis pigmentosa, stroke, degenerative neuropathy, xanthoma or
xanthelasma.
29. The method of claim 28, wherein said degenerative neuropathy is
associated with diabetic neuropathy or multiple sclerosis.
30. A method of elevating high density cholesterol, comprising
administration of a bisphosphonate derivative of the formula
according to claim 20.
31. A method for preventing and/or treating atherosclerosis,
comprising administration of a bisphosphonate derivative of the
formula according to claim 20.
32. A method for preventing and/or treating macular degeneration
and retinitis pigmentosa, comprising administration of a
bsiphosphonate derivative of the formula according to claim 20.
33. A method for the preventing and/or treating stroke, comprising
administration of a bisphosphonate derivative of the formula
according to claim 20.
34. A method for the prevention of degenerative neuropathy,
comprising administration of a bisphosphonate derivative of the
formula according to claim 20.
35. The method of claim 34, wherein said degenerative neuropathy is
associated with diabetic neuropathy or multiple sclerosis.
36. The method of claim 34, wherein said modulation of said apoE
levels in said patient comprises decreasing said apoE levels.
37. The method of claim 36, wherein said patient expresses apoE4,
apoE Leiden or a non-functional mutant form,of apoE.A
38. The method of claim 36, wherein said patient is suffering from
atherosclerosis or Alzheimer's disease.
39. A method for the prevention and/or treatment of Alzheimer's
disease or dementia comprising administration to a patient an
effective amount of bisphosphonate derivative of the formula
according to claim 20.:
40. The method of claim 39, wherein said patient is heterozygous or
homozygous for apoE2 and/or apoE3 and wherein said bisphosphonate
derivative increases apoE levels in said patient.
41. The method of claim 39, wherein said patient is heterozygous or
homozygous for apoE4 and said bisphosphonate derivative decreases
apoE levels in said patient.
Description
FIELD OF INVENTION
[0001] The present invention relates to 1,1- and 1,2-bisphosphonate
compounds, the processes for their preparation, pharmaceutical
compositions containing them and their use in therapy, in
particular for modulating (increasing and decreasing)
apolipoprotein E in plasma and in tissues.
BACKGROUND OF THE INVENTION
[0002] Apolipoprotein E (apoE) is a polymorphic, multifunctional
protein synthesized by several cell types and tissues, including
liver, kidney, skin, adipose tissue, macrophages and brain. The
wide distribution of apoE is associated with the maintenance of key
cellular functions such as intracellular cholesterol trafficking,
cholesterol distribution between cells, and tissue reparation.
[0003] The amino acid sequence of the apoE protein is well
conserved throughout species. ApoE can be viewed as a regulator of
cholesterol homeostasis in tissues such as the central nervous
system (CNS) and peripheral nervous system (PNS) and the arterial
wall (cell-cell) or between tissues via the circulating plasma
lipoproteins (tissue-tissue).
[0004] The major role of plasma apoE containing lipoproteins is to
transfer lipids (cholesterol) from peripheral tissues to the liver
and to remove excess cholesterol from peripheral tissues via the
reverse cholesterol transport system. Dysregulation of this
mechanism leads to excess cholesterol deposition in peripheral
tissues such as arteries (arteriosclerosis) and skin (xanthomas and
xanthelasmas). ApoE has also been shown to have a direct effect on
lymphocyte proliferation and thus has an immunomodulatory role.
[0005] ApoE is the only lipoprotein synthesized in the brain and
has a key role in cholesterol transport between cells of the CNS.
Local secretion of apoE by cells such as macrophages or
macrophage-derived cells is essential for the uptake of excess
tissue cholesterol and the provision of cholesterol for specific
needs such as nerve repair and remyelinisation.
[0006] Up to the present time, compounds affecting Apo E production
in vitro and in vivo have not been extensively investigated. Only
hormone-like estrogens and corticoids have been shown to change Apo
E levels under various experimental conditions (Srivastava et al.,
1997; Stone et al., 1997).
[0007] There is currently a need for compounds that modulate apoE
synthesis and secretion, such compounds having application in the
treatment of diseases such as atherosclerosis, excess lipid
deposition in peripheral tissues such as skin (xanthomas), stroke,
memory loss, optic nerve and retinal pathologies (i.e., macular
degeneration, retinitis pigmentosa), repair of traumatic damage of
the central nervous system (brain tissue), repair of traumatic
damage of the peripheral nervous system (i.e., nerve section
compression or crush), prevention of the degenerative process due
to aging (i.e., Alzheimer's disease), prevention of degenerative
neuropathies occurring in diseases such as diabetic neuropathies
and multiple sclerosis, autoimmune diseases and activation of the
innate immune system.
SUMMARY OF THE INVENTION
[0008] The Applicants have now found that certain 1,1- and
1,2-bisphosphonate compounds modulate (increase or decrease) the
production of apoE in vitro and in vivo. One aspect of the current
invention are bisphosphonate derivatives of formula (I): ##STR1##
wherein Y is hydrogen, hydroxy, halo, aryl, aryloxy,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy or A-L-O; A is
hydroxy, aryl, heterocycle or --NR.sup.3R.sup.4 wherein R.sup.3 and
R.sup.4 are independently hydrogen or C.sub.1-C.sub.4 alkyl. The
bond depicted by represents a single or a double bond. L is
--(CH.sub.2).sub.m--, --(CH.sub.2).sub.pO(CH.sub.2).sub.q--,
--(CH.sub.2).sub.pNR.sup.5(CH.sub.2).sub.q-- or
--(CH.sub.2).sub.pNHCO(CH.sub.2).sub.q--, wherein R.sup.5 is
hydrogen, C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.3 cyanoalkyl; and
subscripts "m," "p" and "q" are an integer from 0 to 6. R.sup.1 and
R.sup.2 are independently hydrogen or C.sub.1-C.sub.6 alkyl. M is
(CH.sub.2).sub.n or (CH.dbd.CH).sub.u--CH.dbd. where n is an
integer from 0 to 3 and u is 0 or 1, with the proviso that if n is
1, 2, or 3 or M is (CH.dbd.CH).sub.u--CH.dbd., then s is 1 and/or
Y, Z.sup.1 and Z.sup.2 are not all independently H, hydroxy, alkyl
or alkoxy. B is H or C.sub.1-C.sub.4 alkyl group and subscript "w"
is 0 when is a double bond and is 1 when is a single bond, it being
understood that the valency of the atoms is respected. Subscript w
is also 1 when M is (CH.sub.2).sub.n and subscript "n" is 0,
wherein there is direct single bond between the substituted phenyl
group and the carbon alpha to the PO(OR.sup.2) group. The subscript
"s" is 0 or 1. Z.sup.1 and Z.sup.2 are independently hydrogen,
C.sub.1-C.sub.4 alkyl, or C.sub.1-C.sub.4 alkoxy.
[0009] The invention also encompasses pharmaceutically acceptable
salts of the compounds of formula (I).
[0010] In various embodiments, n is 0 or 1 and Z.sup.1 and Z.sup.2
are hydrogen. In some embodiments Y is A-O-L-. A may suitably be
pyridin-2-yl, pyridin-3-yl, pyrrolidino, succinimido, piperidino,
morpholino, phthalimido, phenyl, N, N'-(2-cyanoethyl)phenylamino or
p-cyanophenyl. In some embodiments R.sup.1 and R.sup.2 are methyl,
ethyl or isopropyl. In some embodiments the bisphosphonate
derivative of formula (I) is tetraethyl
1-[4-(3-N-phthalimido-propoxy)-phenyl]-methylidene-1,1-bisphosphonate,
tetraethyl
1-[4-(3-N-phthalimido-propoxy)-phenyl]-ethylidene-1,2-bisphosphonate,
tetraethyl
1-{4-[3-(methyl-pyridin-2-yl-amino)-propoxy]-phenyl}-methylidene-1,1-bisp-
hosphonate, or tetraethyl
1-(4-{2-[(2-cyano-ethyl)-phenyl-amino]-ethoxy}-phenyl)-methylidene-1,1-bi-
sphosphonate.
[0011] Other aspects of the current invention include methods of
modulating the production of apoE comprising contacting an apoE
producing cell with an effective amount bisphosphonate derivative
of formula(I) and of modulating apoE levels in a patient in need of
such treatment, comprising administration of an effective amount of
a compound of formula (I). In some embodiments, the levels of apoE
are increased and the patient may be suffering from
atherosclerosis, Alzheimer's disease, macular degeneration,
retinitis pigmentosa, stroke, degenerative neuropathy, xanthoma or
xanthelasma. Increasing apoE levels may provide methods for
elevating high density cholesterol, preventing and/or treating
atherosclerosis, macular degeneration, retinitis pigmentosa, stroke
or degenerative neuropathy. Degenerative neuropathy may be
associated with diabetic neuropathy or multiple sclerosis. In other
embodiments, apoE levels are decreased by administration to a
patient of an effective amount of a bisphosphonate derivative of
formula (I). The patient may express apoE4, apoE Leiden or a
non-functional mutant form of apoE. The patient may be suffering
from atherosclerosis or Alzheimer's disease.
[0012] A further aspect of the invention, provides for a method for
the prevention and/or treatment of Alzheimer's disease or dementia
comprising administration to a patient an effective amount of a
bisphosphonate derivative of formula (I). The patient may be
heterozygous or homozygous for apoE2 and/or apoE3 and the
administration of an effective amount of a bisphosphonate
derivative of formula (I) increases apoE levels. Alternatively, the
patient may be heterozygous or homozygous for apoE4 and the
administration of an effective amount of a bisphosphonate
derivative of formula (I) decreases apoE levels.
BRIEF DESCRIPTION OF THE DRAWINGS
[0013] The following drawings form part of the present
specification and are included to further demonstrate certain
aspects of the present invention. The invention may be better
understood by reference to one or more of these drawings in
combination with the detailed description of specific embodiments
presented herein.
[0014] FIG. 1--Schematic summary of preparation of
methylidene-1,1-diphosphonates of formula (Ia). Substituents Y, A,
L, Z.sup.1, Z.sup.2, R.sup.1 are as described in Detailed
Description of the Invention.
[0015] FIG. 2--Schematic summary of preparation of
alkylidene-1,1-bisphosphonates of formula (Ib) and
alkenylidene-1,1-bisphosphonates of formula (Ic). Substituents Y,
A, L, Z.sup.1, Z.sup.2, R.sup.1 are as described in Detailed
Description of the Invention.
[0016] FIG. 3--Schematic summary of preparation of alkenylidene
phosphonates of formula (Id) and ethylidene-1,2-bisphosphonates of
formula (Ie). Substituents Y, A, L, Z.sup.1, Z.sup.2, R.sup.1 are
as described in Detailed Description of the Invention.
DETAILED DESCRIPTION OF THE INVENTION
I. 1,1- and 1,2-bisphosphonate Compounds
[0017] The present invention relates to 1,1- and 1,2-bisphosphonate
compounds of general formula (I) that modulate (increase or
decrease) apoE levels and are useful as agents for the treatment of
a number of disorders including cardiovascular and neurological
disease states.
[0018] As used herein, the term "aryl" refers to an unsaturated
aromatic carbocyclic group of from 6 to 14 carbon atoms having a
single ring (e.g., phenyl) or multiple condensed (fused) rings
(e.g., naphthyl or anthryl). Suitable aryls include phenyl,
naphthyl and the like. Unless otherwise constrained by the
definition for the aryl substituent, such aryl groups can
optionally be substituted with from 1 to 5 substituents and
preferably 1 to 3 substituents selected from the group consisting
of hydroxy, alkyl (e.g., methyl, ethyl, n-propyl, isopropyl,
sec-butyl, or tert-butyl), alkoxy (e.g., methoxy, ethoxy, propoxy,
tert-butoxy), cyano, amidino, cyanoalkyl (e.g., cyanomethyl,
cyanoethyl and cyanopropyl), aryl, halo e.g., (I, Br, Cl, F) or
nitro.
[0019] As used herein, the term "heterocycle" refers to aromatic
and non-aromatic heterocyclic groups and refers to a single ring or
fused rings containing up to four heteroatoms in at least one ring,
each of which is selected from oxygen, nitrogen and sulphur, which
single or fused ring may be unsubstituted or substituted. Each ring
suitably has from 4 to 7, preferably 5 or 6 ring atoms.
Representative examples of heterocyclic groups include 2-pyridinyl,
3-pyrridinyl, succinimido, pyrrolidino, naphthalimido,
phathalimido, morpholino and pipenridino.
[0020] Pharmaceutically acceptable salts for use in the present
invention include those described by Berge et al. (1997), herein
incorporated by reference. Such salts may be formed from inorganic
and organic acids. Representative examples thereof include maleic,
fumaric, benzoic, ascorbic, pamoic, succinic,
bismethylenesalicylic, methanesulfonic, ethanedisulfonic, acetic,
propionic, tartaric, salicylic, citric, gluconic, aspartic,
stearic, palmitic, itaconic, glycolic, p-aminobenzoic, glutamic,
benzenesulfonic, hydrochloric, hydrobromic, sulfuric,
cyclohexylsulfamic, phosphoric and nitric acids.
[0021] Since the compounds of the present invention, in particular
compounds of formula (I), are intended for use in pharmaceutical
compositions, it will be understood that they are each provided in
substantially pure form, for example at least 50% pure, more
suitably at least 75% pure, preferably at least 95% pure, and more
preferably at least 99% pure (% are on a wt/wt basis). Impure
preparations of the compounds of formula (I) may be used for
preparing the more pure forms used in the pharmaceutical
compositions. Although the purity of intermediate compounds of the
present invention is less critical, it will be readily understood
that the substantially pure form is preferred as for the compounds
of formula (I). Preferably, whenever possible, the compounds of the
present invention are obtained in crystalline form.
[0022] When some of the compounds of this invention are allowed to
crystallise or are recrystallised from organic solvents, solvent of
crystallisation may be present in the crystalline product. This
invention includes within its scope such solvates. Similarly, some
of the compounds of this invention may be crystallised or
recrystallised from solvents containing water. In such cases water
of hydration may be formed. This invention includes within its
scope stoichiometric hydrates as well as compounds containing
variable amounts of water that may be produced by processes such as
lyophilisation. In addition, different crystallisation conditions
may lead to the formation of different polymorphic forms of
crystalline products. This invention includes within its scope all
polymorphic forms of the compounds of formula (I).
II. Applications of ApoE Modulators
[0023] A. ApoE in Atherosclerosis
[0024] As a component of all lipoprotein fractions, apoE plays an
important role in cholesterol homeostasis, by mediating their
interaction with receptors such as the apoB, low-density
lipoprotein (LDL) and other specific receptors. The important role
of apoE in cardiovascular diseases is demonstrated by the apoE
knock-out mouse model, where the animals rapidly develop
hypercholesterolemia and atherosclerosis with pathological features
similar to human atherosclerosis (Plump, 1997). In addition, the
absence of a functional apoE in humans is associated with
abnormally high plasma levels of cholesterol and triglycerides and
the rapid development of atherosclerosis, notwithstanding a low fat
diet (Richard et al., 1995). In the knock-out mouse model, these
changes are prevented by infusion of apoE, transplantation of
macrophage producing apoE, or gene therapy by introducing the human
apoE gene into apoE knock out mice (Linton et al., 1995). These
results indicate a direct beneficial role for apoE and,
consequently, a utility for compounds that increase the apoE
levels. The 1,1- and 1,2-bisphosphonate compounds of the present
invention that increase apoE plasma levels will decrease plasma
atherogenic lipoproteins (VLDL, IDL and LDL) by increasing their
uptake by the liver. Increasing apoE in HDL will increase the
removal of cholesterol from loaded tissues (atherosclerotic
arteries) by the reverse cholesterol transport mechanism.
[0025] In contrast, hyperlipidemic patients susceptible of
developing atherosclerosis due to the expression of a mutated form
of apoE, such as apoE Leiden or other variants, should benefit from
the treatment with the compounds that decrease apoE production (van
Vlijmen et al., 1998; Richard, 1995). Thus, 1,1- and
1,2-bisphosphonate compounds of the present invention that decrease
the production of apoE are useful in the prevention and/or
treatment of pathological cardiovascular conditions secondary to
the presence of non-functional, variants or mutant forms of the
apoE molecule.
[0026] B. ApoE in the Central Nervous System (CNS)
[0027] ApoE also plays a critical role in the CNS. In the brain
apoE is synthesized and secreted by astrocytes, its principal role
being cholesterol transport between cells. ApoE is considered to
redistribute lipids and to participate in the cholesterol
homeostasis of the brain.
[0028] ApoE is linked to the neuropathological lesions
characteristic of Alzheimer's disease. One isoform, ApoE4, is
strongly associated with the age of onset of the disease (Poirier,
1994; Rubinsztein, 1995), while another isoform, apoE3, is believed
to help maintain healthy microtubules. The increase in both apoE
mRNA and the number of astrocytes in the brains of Alzheimer's
patients, indicates that increased apoE represents an astrocyte
repair-mechanism to ameliorate the damage within the nervous cells.
Memory deficit, defective repair of brain injury and deposition of
the Alzheimer's associated .beta.-amyloid variant APPV.sub.717F
have been demonstrated in in the absence of the apoE gene, i.e.,
apoE knock out mice (Oitzl et al., 1997; Laskowitz et al., 1997;
Walker et al., 1997).
[0029] Thus, there is a benefit to increasing apoE production in
patients bearing the E2 and E3 isoforms of apoE in regard to the
occurrence of Alzheimer's or other spontaneous or traumatic
neurological diseases. The 1,1- and 1,2-bisphosphonate compounds of
the present invention that increase apoE in the brain will prevent
the deposition of plaques associated with Alzheimer's disease and
increase the repair mechanism of brain injuries due to mechanical
traumas or strokes. Through the increase of neurite extension
synaptic sprouting the overall brain activity (e.g., memory) should
improve.
[0030] Conversely, patients at risk of or suffering from
Alzheimer's or spontaneous or traumatic neurological diseases who
overexpress the pathological isoforms of apoE, such as apoE4,
should benefit from the treatment with a compound that decreases
apoE. Thus, 1,1- and 1,2-bisphosphonate compounds of the present
invention that decrease the production of apoE are useful in the
prevention and/or treatment of the symptomatic and
neuropathological cardiovascular conditions characteristic of
Alzheimer's or other spontaneous or traumatic neurological diseases
that are caused or exacerbated by non-functional, variants or
mutant forms of the apoE.
[0031] C. ApoE in the Peripheral Nervous System (PNS)
[0032] The important role of apoE in nerve regeneration in the PNS
is demonstrated by the observation that apoE synthesis is
dramatically induced when nerves are injured (Poirier, 1994). The
maintenance and/or repair of the myelin sheets involves the
participation of apoE secreted by support cells such as glial and
Schwann cells. Both apoE synthesis and concentration were found to
be abnormally low in degenerative diseases of nervous tissues such
as in multiple sclerosis (Gaillard, 1996). ApoE is also considered
to stabilize the cytoskeleton apparatus and support neurite
elongation, thus having a major effect on the development and
remodelling following injury of the nervous system occurring late
in life. Thus, the compounds of the present invention that increase
apoE will support and increase the speed of the healing process of
traumatised nerves (nerve section, crush, etc.) and the prevention
and/or healing of degenerative nerves (e.g., multiple
sclerosis).
[0033] D. ApoE as Modulators of the Immune System
[0034] ApoE affects the immune system by acting on lymphocyte
proliferation. Furthermore apoE knock out mice are highly sensitive
to bacterial infection due to a defect in the innate immune system,
suggesting that increasing apoE production should augment the
immune response (Roselaar & Daugherty, 1998). Increasing apoE
production by utilization of compounds of the present invention
should augment ameliorate the immune response in patients in need
thereof
[0035] E. Skin Lipid Metabolism Disorders
[0036] Lipid homeostasis is well controlled in epithelial cells
such as keratinocytes, wherein exported lipids are important for
comeocyte adhesion and for forming the cutaneous barrier to the
external environment. Excess cholesterol deposition in skin
(xanthomas and xanthelasmas) will be prevented by utilization of
1,1- and 1,2-diphosphonates compounds of the present invention that
increase the level of cutaneous apoE.
III. Formulations and Administration
[0037] The compounds of formula (I) can be administered by any of a
variety of routes. Thus, for example, they can be administered
orally, or by delivery across another mucosal surface (for example
across the nasal, buccal, bronchial or rectal mucosa),
transdermally, or by injection (for example intradermal,
intraperitoneal, intravenous or intramuscular injection).
[0038] When the compounds are intended for oral administration,
they can be formulated, for example, as tablets, capsules,
granules, pills, lozenges, powders, solutions, emulsions, syrups,
suspensions, or any other pharmaceutical form suitable for oral
administration. Oral dosage forms can, if desired, be coated with
one or more release delaying coatings to allow the release of the
active compound to be controlled or targeted at a particular part
of the enteric tract.
[0039] Tablets and other solid or liquid oral dosage forms can be
prepared (e.g. in standard fashion) from the compounds of formula
(I) and a pharmaceutically acceptable solubilizer, diluent or
carrier. Examples of solubilizers, diluents or carriers include
sugars such as lactose, starches, cellulose and its derivatives,
powdered tracaganth, malt, gelatin, talc, stearic acid, magnesium
stearate, calcium sulfate, vegetable oils, polyols such as
glycerol, propyleneglycol and polyethyleneglycols, alginic acids
and alginates, agar, pyrogen free water, isotonic saline, phosphate
buffered solutions, and optionally other pharmaceutical excipients
such as disintegrants, lubricants, wetting agents such as sodium
lauryl sulfate, coloring agents, flavoring agents and
preservatives, etc.
[0040] Capsules can be of the hard or soft variety and can contain
the active compound in solid, liquid or semisolid form. Typically
such capsules are formed from gelatine or an equivalent substance
and can be coated or uncoated. If it is desired to delay the
release of the active compound until the capsule has passed through
the stomach and into the intestine, the capsule can be provided
with a pH sensitive coating adapted to dissolve at the pH found in
the duodenum or ileum. Examples of such coatings include the
Eudragits, the uses of which are well known.
[0041] Formulations for injection will usually be made up of the
appropriate solubilizers such as detergents which may also include
compounds and excipients such as buffering agents to provide an
isotonic solution having the correct physiological pH. The
injectable solutions are typically pyrogen-free and can be provided
in scaled vials or ampoules containing a unit dose of compound.
[0042] A unit dosage form of the compounds of the invention
typically will contain from 0.1% to 99% by weight of the active
substance, more usually from 5% to 75% of the active substance. By
way of example, a unit dosage form can contain from lmg to lg of
the compound, more usually from 10 mg to 500 mg, for example
between 50 mg and 400 mg, and typically in doses of 100 mg to 200
mg.
[0043] The compounds of the invention will be administered in
amounts which are effective to provide the desired therapeutic
effect. The concentrations necessary to provide the desired
therapeutic effect will vary according to among other things the
precise nature of the disease, the size, weight and age of the
patient and the severity of the disease. The doses administered
will preferably be non-toxic to the patient, although in certain
circumstances the severity of the disease under treatment may
necessitate administering an amount of compound that causes some
signs of toxicity.
[0044] Typically, the compounds of the invention will be
administered in amounts in the range 0.01 mg/kg to 100 mg/kg body
weight, more preferably 0.1 mg/kg to 10 mg/kg body weight and
particularly 1 mg/kg to 5 mg/kg body weight. For an average human
of 70 kg weight, a typical daily dosage of the compounds of the
invention would be in the range of 70 mg to 700 mg. Such a dosage
can be administered, for example from two to four times daily.
Ultimately however, the size of the doses administered and the
frequency of administration will be at the discretion and judgment
of the physician treating the patient.
[0045] For therapeutic use the compounds of the present invention
will generally be administered in a standard pharmaceutical
composition obtained by admixture with a pharmaceutical carrier
selected with regard to the intended route of administration and
standard pharmaceutical practice. For example, they may be
administered orally in the form of tablets containing such
excipients as starch or lactose, or in capsule, ovules or lozenges
either alone or in admixture with excipients, or in the form of
elixirs or suspensions containing flavoring or coloring agents.
They may be injected parenterally, for example, intravenously,
intramuscularly or subcutaneously. For parenteral administration,
they are best used in the form of a sterile aqueous solution that
may contain other substances, for example, enough salts or glucose
to make the solution isotonic with blood. The choice of form for
administration as well as effective dosages will vary depending,
inter alia, on the condition being treated. The choice of mode of
administration and dosage is within the skill of the art.
[0046] The compounds of formula (I) and their pharmaceutically
acceptable salts which are active when given orally can be
formulated as liquids, for example syrups, suspensions or emulsions
or as solids for example, tablets, capsules and lozenges. A liquid
formulation will generally consist of a suspension or solution of
the compound or pharmaceutically acceptable salt in suitable liquid
carrier(s) for example, ethanol, glycerine, non-aqueous solvent,
for example polyethylene glycol, oils, or water with a suspending
agent, preservative, flavoring or coloring agents. A composition in
the form of a tablet can be prepared using any suitable
pharmaceutical carrier(s) routinely used for preparing solid
formulations. Examples of such carriers include magnesium stearate,
starch, lactose, sucrose and cellulose. A composition in the form
of a capsule can be prepared using routine encapsulation
procedures. For example, pellets containing the active ingredient
can be prepared using standard carriers and then filled into a hard
gelatin capsule; alternatively, a dispersion or suspension can be
prepared using any suitable pharmaceutical carrier(s), for example
aqueous gums, celluloses, silicates or oils and the dispersion or
suspension then filled into a soft gelatine capsule.
[0047] Typical parenteral compositions consist of a solution or
suspension of the compound or pharmaceutically acceptable salt in a
sterile aqueous carrier or parenterally acceptable oil, for example
polyethylene glycol, polyvinyl pyrrolidone, lecithin, arachis oil
or sesame oil. Alternatively, the solution can be lyophilised and
then reconstituted with a suitable solvent just prior to
administration.
[0048] A typical suppository formulation comprises a compound of
formula (I) or a pharmaceutically acceptable salt thereof which is
active when administered in this way, with a binding and/or
lubricating agent such as polymeric glycols, gelatins or cocoa
butter or other low melting vegetable or synthetic waxes or
fats.
[0049] Preferably the composition is in unit dose form such as a
tablet or capsule.
[0050] Each dosage unit for oral administration contains preferably
from 1 to 250 mg (and for parenteral administration contains
preferably from 0.1 to 25 mg) of a compound of the structure (I) or
a pharmaceutically acceptable salt thereof calculated as the free
base.
[0051] The pharmaceutically acceptable compounds of the invention
will normally be administered to a subject in a daily dosage
regimen. For an adult patient this may be, for example, an oral
dose of between 1 mg and 500 mg, preferably between I mg and 250
mg, or an intravenous, subcutaneous, or intramuscular dose of
between 0.1 mg and 100 mg, preferably between 0.1 mg and 25 mg, of
the compound of the structure (I) or a pharmaceutically acceptable
salt thereof calculated as the free base, the compound being
administered I to 4 times per day.
[0052] Disease states which could benefit from increasing plasma
and tissue apoE levels include, but are not limited to:
atherosclerosis, neurodegenerative disorders such as Alzheimer's
disease or dementia. The compounds of this invention modulate apoE
and are therefore of value in the treatment of any of these
conditions.
[0053] Compounds of the present invention may also be of use in
preventing and/or treating the above mentioned disease states in
combination with anti-hyperlipidaemic, anti-atherosclerotic,
anti-diabetic, anti-anginal, anti-inflammatory or anti-hypertension
agents. Examples of the above include cholesterol synthesis
inhibitors such as statins, for instance atorvastatin, simvastatin,
pravastatin, cerivastatin, fluvastatin, lovastatin and ZD 4522
(also referred to as S-4522, Astra Zeneca), anti-oxidants such as
probucol, insulin sensitisers such as a PPAR gamma activator, for
instance G1262570 (Glaxo Wellcome) and the glitazone class of
compounds such as rosiglitazone (Avandia, SmithKline Beecham),
troglitazone and pioglitazone, calcium channel antagonists, and
anti-inflammatory drugs such as NSAIDs.
IV. Synthesis of Bisphosphonate Compounds of Formula (I)
[0054] The present invention also provides methods for the
preparation of compounds of formula (I). Such methods include the
preparation of methylidene-1,1-bisphosphonates of formula (Ia):
##STR2## alkylidene-1,1-bisphosphonates of formula (Ib): ##STR3##
alkenylidene-1,1-bisphosphonates of formula (Ic): ##STR4##
alkenylidene phosphonates of formula (Id): ##STR5## and
ethylidene-1,2-bisphosphonates of formula (Ie): ##STR6## where n,
u, Y, Z.sup.1, Z.sup.2 , R.sup.1 and R.sup.2 are as described
previously.
[0055] The first method, shown schematically in FIG. 1, provides
for the preparation of methylene-1,1-bisphosphonates wherein
R.sup.1 is the same as R.sup.2. The experimental procedure consists
of the derivatization of the phenol-methylene-1,1-bisphosphonate
intermediate (IV) obtained by reacting the hydroxybenzaldehyde (II)
with the dialkyl phosphite (III) in presence of an amine such as
diisopropylamine. The methylene-1,1-bisphosphonate (Ia) is prepared
from the phenol-methylene-1,1-bisphosphonate (IV) by means of a
Mitsunobu or a Williamson reaction. In the Mitsunobu reaction the
phenol bisphosphonate (IV) is reacted with the primary alcohol (V),
wherein X is OH, in presence of a mixture of dialkyl
azodicarboxylate and triphenylphosphine. In the Williamson reaction
the phenol bisphosphonate (IV) is reacted with the alkyl halide
(V), wherein X is a halide, in presence of a base.
[0056] The second method, outlined in FIG. 2, provides for the
preparation of alkenylidene-1,1-bisphosphonates of formula (Ic) and
alkylidene-1,1-bisphosphonates of formula (Ib). One experimental
procedure consists in the derivatization of the
phenol-alkylidene-1,1-bisphosphonate (VIII). Compound (VII) is
obtained by reacting the hydroxybenzaldehyde (II) with the
tetraalkyl methylenebisphosphonate (VI) in presence of titanium
tetrachloride and a tertiary amine such as methylmorpholine or
pyridine. Reduction of (VII) by a complex hydride reagent such as
sodium borohydride or lithium borohydride or by catalytic
hydrogenation gives the phenol-alkylidene-1,1-bisphosphonate
compound (VIII). Suitable methods of derivatization of compound
(VIII) are the Mitsunobu or Williamson reactions. In the Mitsunobu
reaction, the phenol bisphosphonate (VIII) is reacted with the
primary alcohol (V), wherein X is OH, in presence of a mixture of
dialkyl azodicarboxylate and triphenylphosphine. In the Williamson
reaction, the phenol bisphosphonates (VIII) is reacted with the
alkyl halide (V), wherein X is a halide, in presence of a base.
Alternatively, the compounds of formula (Ib) can be prepared by
reacting the already substituted hydroxybenzaldehyde (IX) with
tetraalkyl methylenebisphosphonate (VI), titanium tetrachloride and
methylmorpholine.
[0057] An alternative method of preparing (Ic) consists of
derivatizing compound (VII) by a Mitsunobu or a Williamson reaction
in the conditions described above for compounds of formula
(Ib).
[0058] Alkenylidene phosphonates of formula (Id) and
ethylidene-1,2-bisphosphonates of formula (Ie) may be prepared as
shown in FIG. 3. The protected hydroxybenzaldehyde (X) is reacted
with the tetraalkyl methylenebisphosphonate (XI) under
Horner-Emmons conditions to give the vinylphosphonate (XII).
Addition of dialkyl phosphite (III) over sodium hydride provides
the protected phenol-1,2-bisphosphonate (XIV). Deprotection of
(XIV) provides the free phenol (XV), which is then converted into a
compound of formula (Ie) by means of the Mitsunobu reaction or the
Williamson reaction as described above. The alkenylidene
phosphonates (Id) is prepared by means of a Mitsunobu or a
Williamson reaction from the free phenol (XIII) obtained by
deprotection of the vinylphosphonate (XII).
[0059] Alternatively, compounds (Id) and (Ie) can be prepared by
reacting the already substituted hydroxybenzaldehyde (XVI) with
tetraalkyl methylenebisphosphonate (XI) to give the substituted
alkenylidene phosphonate (Id) and reacting this with the dialkyl
phosphite (XIV) and sodium hydride.
V. Determination of Biological Activity
[0060] The bisphosphonate compounds of the invention can modulate
(increase or decrease) apoE levels in plasma and in tissues. The
activities of the compounds can be determined in an in vitro cell
assay comprising determining the modulatory effect of the test
compound on the secretion of apoE by an apoE secreting cell line
(e.g., a monocyte-macrophage cell line such as the THP-1 cell line,
a liver derived cell line such as the HepG2 cell line, an
intestinal derived cell line such as the CaCo2 cell line or a brain
derived cell line such as the astrocytoma CCF-STTG1 cell line).
EXAMPLES OF THE INVENTION
[0061] Without further elaboration, it is believed that one skilled
in the art can, using the preceding description, utilize the
present invention to its fullest extent. The following specific
examples are intended merely to illustrate the invention and not to
limit the scope of the disclosure or the scope of the claims in any
way whatsoever.
[0062] All new products were purified by flash chromatography
employing silica gel 60 Fluka 60752 or standard chromatography with
silica gel from Chemie Brunschwig CB 09363-25; analytical TLC on
silica gel 60 F.sub.254 aluminium sheets from Merck. Detection by
UV light at 254 nm. The purity of all new products was determined
by GC (HP6890 or HP5890, optima5, t.sub.R in min.).
[0063] NMR spectra were performed using a Bruker AMX400 (.sup.1H at
400 MHz) or Bruker AMX-500 (.sup.1H at 500 MHz). Chemical shifts 8
in ppm with respect to SiMe.sub.4 (.delta.=0 ppm, internal
reference). For .sup.31P-NMR, chemical shifts o are in ppm with
respect to H.sub.3PO.sub.4 (.delta.=0 ppm, external reference 85%
H.sub.3PO.sub.4 in H.sub.2O). Coupling constants are given in Hz.
Nonobvious signal assignment were made by comparison with the
spectra of the described similar compounds.
[0064] MS were carried out with a Varian CH4 or SM1 spectrometer
with electron impact or electrospray, m/z (% of base peak).
[0065] IR spectra were carried out with a Perkin Elmer Paragon 1000
FT-IR spectrometer. KBr for solids and neat for oil or liquids.
Absorption bands in cm.sup.-1.
[0066] All reactions were conducted under N.sub.2. Reaction
temperatures refer to that of the heating bath. Reactions carried
out with the exclusion of light were performed in flasks completely
wrapped in aluminium foil. All reactions were monitored by TLC
and/or GC upon total consumption of the starting material.
Example 1
Tetramethyl 1-(4-hydroxy-phenyl)methylidene-1,1-bisphosphonate
[0067] ##STR7##
[0068] Diisopropyl amine (24 ml, 169.82 mmol) was added to a
stirred mixture of 20.22 g (165.58 mmol) of 4-hydroxybenzaldehyde
and 76.0 ml (829.4 mmol) of dimethyl phosphite. The reaction
mixture was heated at 110.degree. C. for 5 h, then the excess of
phosphite and amine were removed under vacuum. The mixture was
poured into water (100 ml) and taken up with CH.sub.2Cl.sub.2
(6.times.50 ml). The organic layer was washed with water, saturated
NaCl solution and dried over MgSO.sub.4. The solvent was evaporated
by rotary evaporator under reduced pressure. The oily residue
(79.05 g) was purified by silica gel chromatography (AcOEt:MeOH
80:20). Two products were isolated, the wanted 1,1-bisphosphonate
(C.sub.11H.sub.18O.sub.7P.sub.2, M.sub.w=324.21, R.sub.f 0.24,
33.49 g, 103.30 mmol, yield 41.66%) and a monophosphonate
(C.sub.10H.sub.15O.sub.5P, M.sub.w=246.20, R.sub.f 0.49, 29.18 g,
118.50 mmol, yield 47.80%). The monophosphonate was identified as
dimethyl
1-(4-hydroxy-phenyl)-1-methoxy-methylidene-1-phosphonate.
[0069] .sup.1H-NMR (DMSO, 400 MHz)
[0070] .delta.=9.4, s br, 1H; .delta.=7.26, dxt, 2H, J=8.8, J=2.0;
.delta.=6.69, d, J=8.4; .delta.=4.29, t, 1H, J=25.2; .delta.=3.62,
d, 3H, J=13.6; .delta.=3.45, d, 3H, J=13.2.
[0071] MS (70 eV)
[0072] 325 (11), 324 (M.sup.+, 82), 231 (7), 230 (18), 218 (18),
216 (12), 215 (50), 187 (8), 183 (9), 137 (7), 121 (14), 120 (13),
109 (11), 107 (32), 105 (7), 93 (100), 81 (11), 79 (12), 66 (15),
65 (23), 63 (16), 51 (8), 47 (24).
Example 2
Tetraethyl 1-(4-hydroxy-phenyl)-methylidene-1,1-bisphosphonate
[0073] ##STR8##
[0074] Diisopropyl amine (36 ml, 254.01 mmol) was added to a
stirred mixture of 30.18 g (247.11 mmol) of 4-hydroxybenzaldehyde
and 105.0 ml (815.05 mmol) of diethyl phosphite. The reaction
mixture was heated at 110.degree. C. for 2 h, then the excess of
phosphite and amine were removed under vacuum. The mixture was
poured into water (100 ml) and taken up with CH.sub.2Cl.sub.2
(4.times.100 ml). The organic layer was washed with water,
saturated NaCl solution and dried over MgSO.sub.4. The solvent was
evaporated by rotary evaporator under reduced pressure. The oily
residue (83.57 g) was purified by repeated flash chromatography
(Silica gel 60, AcOEt:MeOH 90:10). Two products were isolated, the
wanted 1,1-bisphosphonate (C.sub.15H.sub.26O.sub.7P.sub.2,
M.sub.w=380.32, R.sub.f 0.25, 56.18 g, 147.72 mmol, yield 59.78%)
and a monophosphonate (C.sub.13H.sub.21O.sub.5P, M.sub.w=288.28,
R.sub.f 0.56, 4.40 g, 15.26 mmol, yield 6.18%). The monophosphonate
was identified as diethyl
1-(4-hydroxy-phenyl)-1-ethoxy-methylidene-1-phosphonate.
[0075] .sup.1H-NMR (CDCl.sub.3, 400 MHz)
[0076] .delta.=8.35, s br, 1H; .delta.=7.22, dxt, 2H, J=8.4, J=2.0;
.delta.=6.66, d, 2H, J=8.4; .delta.=4.19-3.84, m, 4.times.2H;
.delta.=3.64, t, 1H, J=25.2; .delta.=1.31, t, 2.times.3H, J=7.2;
.delta.=1.16, t, 2.times.3H, J=7.2.
[0077] MS (70 eV)
[0078] 381 (27), 380 (M.sup.+, 67), 272 (7), 268 (7), 260 (23), 244
(32), 243 (9), 216 (9), 215 (16), 198 (14), 187 (20), 170 (22), 155
(20), 133 (8), 127 (8), 123 (18), 121 (31), 108 (8), 107 (100), 106
(16), 105 (10), 93 (9), 80 (13), 78 (10), 77 (16), 65 (22).
Example 3
Tetraisopropyl
1-(4-hydroxy-phenyl)-methylidene-1,1-bisphosphonate
[0079] ##STR9##
[0080] Diisopropyl amine (4.28 g, 42.29 mmol) was added to a
stirred mixture of 5.16 g (42.25 mmol) of 4-hydroxybenzaldehyde and
35.13 g (42.29 mmol) of diisopropyl phosphite. The reaction mixture
was heated at 110.degree. C. for 2 h, then the excess of phosphite
and amine were removed under vacuum. The mixture was poured into
water (30 ml) and taken up with CH.sub.2Cl.sub.2 (4.times.30 ml).
The organic layer was washed with water, saturated NaCl solution
and dried over MgSO.sub.4. The solvent was evaporated by rotary
evaporator under reduced pressure. The oily residue was purified by
flash chromatography (Silica gel 60, AcOEt:MeOH 95:5, R.sub.f 0.28)
to give the wanted 1,1-bisphosphonate
(C.sub.19H.sub.34O.sub.7P.sub.2, M.sub.w=436.43, 7.66 g, 17.55
mmol, yield 41.54%).
[0081] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0082] .delta.=5.6-5.3, s br, 1H; .delta.=7.20, d, 2H, J=8.4;
.delta.=6.63, d, 2H, J=8.4; .delta.=4.72, sept, 2.times.1H, J=1.72;
.delta.=4.53, sept, 2.times.1H, J=7.0; .delta.=3.53, t, 1H,
J=25.48; .epsilon.=1.35-1.30, m, 4.times.3H; .delta.=1.27, d,
2.times.3H, J=6.2; .delta.=1.03, d, 2.times.3H, J=6.2.
[0083] MS (70 eV)
[0084] 436 (M.sup.+, 15), 310 (8), 272 (20), 269 (11), 268 (100),
251 (14), 230 (13), 188 (55), 187 (33), 170 (13), 135 (9), 125 (9),
123 (22), 121 (8), 109 (21), 107 (41), 106 (10), 99 (11), 83 (26),
77 (89), 45 (18).
Example 4
Tetraethyl
1-[4(3-hydroxy-propoxy)-phenyl]-methylidene-1,1-bisphosphonate
[0085] ##STR10##
[0086] A solution of 1.3 ml (14.95 mmol) of 3-bromo-1-propanol in
20 ml 2-butanone was added dropwise to a stirred mixture of 4.98 g
(13.09 mmol) of tetraethyl
1-(4-hydroxy-phenyl)-methylidene-1,1-bisphosphonate, 2.77 g (20.03
mmol) of potassium carbonate and 0.05 g (0.144 mmol) of tetrabutyl
ammonium bromide in 40 ml of the same solvent. The reaction mixture
was warmed under reflux overnight then allowed to cool to room
temperature. The final mixture was poured into water and taken up
with CH.sub.2Cl.sub.2 (3.times.20 ml). The organic layer was washed
with saturated NaCl solution, dried over MgSO.sub.4, filtered and
then concentred by rotary evaporator. The residue (6.42 g) was
purified by flash chromatography (Silica gel 60, AcOEt:MeOH 90:10,
R.sub.f 0.31) to give 4.91 g (11.20 mmol,
C.sub.18H.sub.32O.sub.8P.sub.2, M.sub.W=438.40, yield 85.56%) of
tetraethyl
1-[4-(3-hydroxy-propoxy)-phenyl]-methylidene-1,1-bisphosphonate.
[0087] .sup.1H-NMR (CDCl.sub.3, 400 MHz)
[0088] .delta.=7.39, dxt, 2H, J=8.0, J=2.0; .delta.=6.87, d, 2H,
J=8.0; .delta.=4.16-3.82, m, 6.times.2H; .delta.=3.67, t, 1H,
J=25.0; .delta.=2.04, quint, 2H, J=6.0; .delta.=1.29, t,
2.times.3H, J=7.0; .delta.=1.17, t, 2.times.3H, J=7.0.
[0089] MS (70 eV)
[0090] 439 (10), 438 (M.sup.+, 60), 393 (7), 215 (15), 214 (8), 381
(17), 380 (100), 302 (22), 301 (44), 187 (26), 179 (28), 169 (10),
165 (26), 133 (8), 273 (18), 256 (18), 231 (10), 228 (14), 227
(13), 121 (11), 107 (34).
[0091] IR
[0092] 3400 (s br), 2982 (s), 1740 (m), 1610 (m), 1512 (m), 1249
(s), 1045 (s), 875 (m).
Example 5
Tetraethyl
1-{4-[3-N-(1,8-naphthalimido)propoxy]-phenyl}-methylidene-1,1-b-
isphosphonate
[0093] ##STR11##
[0094] Under nitrogen and with exclusion of light, a solution of
1.2 ml (5.93 mmol) of diisopropyl azodicarboxylate and 2.00 g (4.56
mmol) of tetraethyl
1-[4-(3-hydroxy-propoxy)-phenyl]-methylidene-1,1-bisphosphonate in
30 ml CH.sub.2Cl.sub.2 was added very slowly (1 drop/2 sec) to the
mixture of 1.56 g (5.95 mmol) of triphenyl phosphine and 1.10 g
(5.48 mmol) of 1,8-naphthalimide in 40 ml CH.sub.2Cl.sub.2 at room
temperature. The mixture was stirred at room temperature overnight.
After solvent removal by rotary evaporator, the residue was
purified by flash chromatography (Silica gel 60, AcOEt:MeOH 90:10,
R.sub.f 0.27) to give 1.1 g (1.73 mmol,
C.sub.30H.sub.37NO.sub.9P.sub.2, M.sub.W=617.58, yield 37.94%) of
naphthalimido substituted 1,1-bisphosphonate.
[0095] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0096] .delta.=8.61, dxd, 2H, J=7.3, J=1.0; .delta.=8.23, dxd, 2H,
J=8.4, J=1.0; .delta.=7.77, dxd, 2H, J=8.1, J=7.3; .delta.=7.34,
dxt, 2H, J=8.7, J=2.0; .delta.=6.80, d, 2H, J=8.7; .delta.=4.41, t,
2H, J=7.0; .delta.=4.15-3.88, m, 5.times.2H; .delta.=3.66, t, 1H,
J=25.2; .delta.=2.26, quint, 2H, J=7.0; .delta.=1.28, t,
2.times.3H, J=7.1; .delta.=1.16, t, 2.times.3H, J=7.1.
[0097] MS (70 eV)
[0098] 618 (4), 617 (M.sup.+, 7), 393 (15), 380 (5), 245 (7), 239
(51), 238 (100), 211 (8), 210 (47), 203 (10), 187 (5), 180 (10),
161 (9), 107 (7).
Example 6
Tetraethyl
1-[4-(3-N-phthalimido-propoxy)-phenyl]-methylidene-1,1-bisphosp-
honate
[0099] ##STR12##
[0100] A solution of 1.67 g (6.23 mmol) of
N-(3-bromopropyl)phthalimide in 10 ml 2-butanone was added dropwise
to a stirred mixture of 2.11 g (5.55 mmol) of tetraethyl
1-(4-hydroxy-phenyl)-methylidene-1,1-bisphosphonate, 1.20 g (8.68
mmol) of potassium carbonate and 0.19 g (0.57 mmol) of tetrabutyl
ammonium bromide in 20 ml of the same solvent. The reaction mixture
was warmed under reflux over 4 h then allowed to cool to room
temperature. The final mixture was poured into water and taken up
with CH.sub.2Cl.sub.2 (3.times.30 ml). The organic layer was washed
with saturated NaCl solution, dried over MgSO.sub.4, filtered and
then concentred by rotavapory evaporator. The residue (3.54 g) was
purified by flash chromatography (Silica gel 60, AcOEt:MeOH 90:10,
R.sub.f 0.27) to give 3.13 g (5.52 mmol,
C.sub.26H.sub.35NO.sub.9P.sub.2, M.sub.W=567.52, oil, yield 94.46%)
of
1-[4-(3-N-phthalimido-propoxy)-phenyl]-methylidene-1,1-bisphosphonate.
[0101] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0102] .delta.=7.84, m, 2H, .delta.=7.72, m, 2H; .delta.=7.34 txd,
2H, J=8.7, J=1.9; .delta.=6.77, d, J=8.7; .delta.=4.15-3.89, m,
6.times.2H; .delta.=3.66, t, 1H, J=25.2; .delta.=2.18, quint, 2H,
J=6.6; .delta.=1.28, t, 3.times.3H, J=7.1; .delta.=1.16, t,
2.times.3H, J=7.1.
[0103] .sup.13C-NMR (CDCl.sub.3, 125 MHz)
[0104] .delta.=168.4, (C); .delta.=158.28, (C); .delta.=133.9,
(CH); .delta.=132.2, (C); .delta.=131.5, (CH), t, J=6;
.delta.=123.2, (CH); .delta.=122.0, (C), t, J=8; .delta.=114.5,
(CH); .delta.=65.7, (CH.sub.2); .delta.=63.1, (CH.sub.2), d, J=75;
.delta.=44.7, (CH), t, J=133; .delta.=35.5, (CH.sub.2);
.delta.=28.3, (CH.sub.2); .delta.=16.3, (CH.sub.3), d, J=11.
[0105] MS (70 eV)
[0106] 568 (1), 567 (M.sup.+, 4), 380 (5), 189 (12), 188 (100), 160
(21), 107 (16), 65 (7).
[0107] IR
[0108] 2983 (s), 1770(w), 1713(s), 1611 (m), 1582 (w), 1513 (w),
1469 (w), 1392 (m), 1234(s), 1662 (m), 1045 (s br), 873 (m), 796
(m), 724 (m), 635 (w), 604 (w), 550 (s), 497 (m).
Example 7
1-[4-(3-N-Phthalimido-propoxy)-phenyl]-methylidene-1,1-bisphosphonic
acid
[0109] ##STR13##
[0110] Trimethylsilyl bromide (5.4 ml, 42.3 mmol) was added to a
solution of 2.40 g (4.23 mmol) of
1-[4-(3-N-phthalimido-propoxy)-phenyl]-methylidene-l,l-bisphosphonate
in 15 ml CCl.sub.4 at -10.degree. C. The mixture was allowed to
warm to room temperature and stirred for 6 h. The solvent and the
excess of trimethylsilyl bromide were removed under vacuum. The
residue was taken up with warm CH.sub.2Cl.sub.2 and the unsoluble
bisphosphonic acid was isolated by filtration. The solid was washed
with warm CH.sub.2Cl.sub.2. We obtained 1.40 g (3.08 mmol,
C.sub.18H.sub.19NO.sub.9P.sub.2; M.sub.W=455.30, white solid,
mp>250.degree. C., yield 72.81%) of
1-[4-(3-N-phthalimido-propoxy)-phenyl)-methylidene-1,1-bisphosphonic
acid.
[0111] .sup.1H-NMR (DMSO, 500 MHz)
[0112] .delta.=9.4-7.9, s br, 4.times.1H; .delta.=7.88-7.82, m,
4.times.1H; .delta.=7.33, d, 2H, J=8.6; .delta.=6.73, d, 2H, J=8.6;
.delta.=3.98, t, 2H, J=5.8; .delta.=3.76, t, 2H, J=6.8;
.delta.=3.48, t, 1H, J=24.6; .delta.=2.05; quint, 2H, J=6.2.
[0113] MS (70 eV)
[0114] 188 (17), 187 (100), 169 (45), 160 (29), 158 (10), 130 (21),
105 (11), 104 (21), 77 (15), 76 (33), 51 (13), 50 (18).
Example 8
Tetraisopropyl
1-[4(3-N-phthalimidopropoxy)-phenyl]-methyliden-1,1-bisphosphonate
[0115] ##STR14##
[0116] A solution of 0.7 g (5.04 mmol) of 3-bromo-1-propanol in
10mi 2-butanone was added dropwise to a stirred mixture of 2.0 g
(4.58 mmol) of tetraisopropyl
1-(4-hydroxy-phenyl)-methylidene-1,1-bisphosphonate, 0.95 g (6.87
mmol) of potassium carbonate and 0.22 g (0.68 mmol) of tetrabutyl
ammonium bromide in 20 ml of the same solvent. The reaction mixture
was warmed under reflux over 4 h then allowed to cool to room
temperature. The final mixture was poured into water and taken up
with CH.sub.2Cl.sub.2 (3.times.20 ml). The organic layer was washed
with saturated NaCl solution, dried over MgSO.sub.4, filtered and
then concentred by rotavapory evaporator.
[0117] The crude tetraisopropyl
1-[4-(3-hydroxy-propoxy)-phenyl]-methylidene-1,1-bisphosphonate
(yield.about.97%) was used in the next step without further
purification. Under nitrogen and with exclusion of light, a
solution of 0.9 ml (5.8 mmol) of diethyl azodicarboxylate and
tetraisopropyl
1-[4-(3-hydroxy-propoxy)-phenyl]-methylidene-1,1-bisphosphonate
(.ltoreq.4.58 mmol) in 30 ml CH.sub.2Cl.sub.2 was added very slowly
(1 drop/2 sec) to the mixture of 1.5 g (5.79 mmol) of triphenyl
phosphine and 0.8 g (5.34 mmol) of phthalimide in 40 ml
CH.sub.2Cl.sub.2 at room temperature. The mixture was stirred at
room temperature over 4 h. After solvent removal by rotary
evaporator, the residue was purified by flash chromatography
(Silica gel 60, AcOEt:MeOH 98:2, R.sub.f 0.18) to give 1.2 g (1.92
mmol, C.sub.30H.sub.43NO.sub.9P.sub.2, M.sub.W=623.63, oil, yield
41.92%) of phthalimido substituted 1,1-bisphosphonate.
[0118] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0119] .delta.=7.85-7.72, m, 2H; .delta.=7.74-7.72, m, 2H;
.delta.=7.32, dxt, 2H, J=8.6, J.times.1.8; .delta.=6.74, d, 2H,
J=8.7; .delta.=4.70, sept, 2.times.1H, J=6.2; .delta.=4.53, sept,
2.times.1H, J=6.3; .delta.=4.02, t, 2H, J=6.1; .delta.=3.91, t, 2H,
J=6.7; .delta.=3.54, t, 1H, J=25.4; .delta.=2.18, quint, 2H, J=6.5;
.delta.=1.30, d, 2.times.3H, J=6.2; .delta.=1.27, d, 2.times.3H,
J=6.2; .delta.=1.25, d, 2.times.3H, J=6.4; .delta.=1.00, d,
2.times.3H, J=6.2.
[0120] MS (70 eV)
[0121] 624 (1), 623 (M.sup.+, 4), 459 (6), 375 (10), 189 (13), 188
(100), 187 (6), 160 (20).
Example 9
Tetraethyl
1-[3-methoxy-5-methyl-4-(3-N-phthalimido-propoxy)-phenyl]-methy-
lidene-1,1-bisphosphonate
[0122] ##STR15##
[0123] A solution of 1.56 g (5.82 mmol) of
N-(3-bromopropyl)phthalimide in 10 ml 2-butanone was added dropwise
to a stirred mixture of 2.06 g (4.85 mmol) of tetraethyl
1-(3-methoxy-5-methyl-4-hydroxy-phenyl)-methylidene-1,1-bisphosphonate,
1.0 g (7.28 mmol) of potassium carbonate and 0.15 g (0.45 mmol) of
tetrabutyl ammonium bromide in 20 ml of the same solvent. The
reaction mixture was warmed under reflux over 4 h then allowed to
cool to room temperature. The final mixture was poured into water
and taken up with CH.sub.2Cl.sub.2 (3.times.30 ml). The organic
layer was washed with saturated NaCl solution, dried over
MgSO.sub.4, filtered and then concentred by rotary evaporator. The
residue (3.2 g) was purified by flash chromatography (Silica gel
60, AcOEt:MeOH 90:10, R.sub.f 0.45) to give 2.0 g (3.27 mmol,
C.sub.28H.sub.39NO.sub.10P.sub.2, M.sub.W=611.57, oil, yield
67.42%) of
1-[4-(3-N-phthalimido-propoxy)-phenyl)-methylidene-1,1-bisphosphonate.
[0124] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0125] .delta.=7.87-7.85, m, 2H; .delta.=7.73-7.71, m, 2H;
.delta.=6.97, s, 1H; .delta.=4.17-3.90, m, 6.times.2H;
.delta.=3.81, s, 3H; .delta.=3.61, t, 1H, J=25.1; .delta.=2.25, s,
3H; .delta.=2.15, quint, 2H, J=7.2; .delta.=1.30, t, 2.times.3H,
J=7.1; .delta.=1.17, t, 2.times.3H, J=7.1.
[0126] MS (70 eV)
[0127] 612 (2), 611 (M.sup.+, 4), 189 (35), 188 (100), 161 (6), 160
(51), 151 (9), 130 (9), 77 (5), 65 (5).
Example 10
Tetraethyl
1-[4-(3-N-phthalimido-butoxy)-phenyl]-methylidene-1,1-bisphosph-
onate
[0128] ##STR16##
[0129] A solution of 1.78 g (6.31 mmol) of
N-(3-bromobutyl)phthalimide in 10 ml 2-butanone was added dropwise
to a stirred mixture of 2.0 g (5.26 mmol) of tetraethyl
1-(4-hydroxy-phenyl)-methylidene-1,1-bisphosphonate, 1.1 g (7.89
mmol) of potassium carbonate and 0.17 g (0.51 mmol) of tetrabutyl
ammonium bromide in 20 ml of the same solvent. The reaction mixture
was warmed under reflux over 4 h then allowed to cool to room
temperature. The final mixture was poured into water and taken up
with CH.sub.2Cl.sub.2 (3.times.30 ml). The organic layer was washed
with saturated NaCl solution, dried over MgSO.sub.4, filtered and
then concentred by rotary evaporator. The residue (3.54 g) was
purified by flash chromatography (Silica gel 60, AcOEt:MeOH 90:10,
is R.sub.f 0.31) to give 2.45 g (4.21 mmol,
C.sub.27H.sub.37NO.sub.9P.sub.2, M.sub.W=581.54, oil, yield 80.04%)
of
1-[4-(3-N-phthalimido-butoxy)-phenyl]-methylidene-1,1-bisphosphonate.
[0130] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0131] .delta.=7.86-7.84, m, 2H; .delta.=7.73-7.72, m, 2H;
.delta.=7.36, dxt, 2H, J=8.7; J=1.8; .delta.=6.84, d, 2H, J=8.7;
.delta.=4.16-4.01 and 3.96-3.89, m, 4.times.2H; .delta.=3.98, t,
2H, J=5.8; .delta.=3.74, t, 2H, J=7.0; .delta.=3.67, t, 1H, J=25.2;
.delta.=1.92-1.81, m, 2.times.2H; .delta.=1.28, t, 2.times.3H,
J=7.1; .delta.=1.16, t, 2.times.3H, J=7.1.
[0132] MS (70 eV)
[0133] 582 (6), 581 (M.sup.+, 16), 393 (5), 203 (15), 202 (100),
161 (9), 160 (78), 130 (8), 107 (16), 105 (5), 77 (6), 65 (6), 55
(8).
Example 11
Tetraethyl
1-(4-{2-[(2-cyano-ethyl)-phenyl-amino]-ethoxy}-phenyl)-methylid-
ene-1,1-bisphosphonate
[0134] ##STR17##
[0135] Under nitrogen and with exclusion of light, a solution of
1.1 ml (6.84 mmol) of diethyl azodicarboxylate and 1.2 g (6.31
mmol) of N-(2-cyanoethyl)-N-(2-hydroxyethyl)-aniline in 40 ml
CH.sub.2Cl.sub.2 was added very slowly (1 drop/2 sec) to the
mixture of 1.8 g (6.84 mmol) of triphenyl phosphine and 2.0 g (5.26
mmol) of tetraethyl
1-(4-hydroxy-phenyl)-methylidene-1,1-bisphosphonate in 40 ml
CH.sub.2Cl.sub.2 at room temperature. The mixture was stirred at
room temperature overnight. After solvent removal by rotary
evaporator, the residue was purified by flash chromatography
(Silica gel 60, AcOEt:MeOH 95:5, R.sub.f 0.20) to give 2.5 g (4.53
mmol, C.sub.26H.sub.38N.sub.2O.sub.7P.sub.2, M.sub.W=552.28, oil,
yield 86.12%) of tetraethyl
1-(4-{2-[(2-cyano-ethyl)-phenyl-amino]-ethoxy}-phenyl)-methylidene-1,1-bi-
sphosphonate.
[0136] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0137] .delta.=7.38, dxt, 2H, J=8.7, J=2.0; .delta.=7.30-7.27, m,
2H; .delta.=6.84, d, 2H, J=8.7; .delta.=6.80, t, 1H, J=7.3;
.delta.=6.72, d, 2H, J=8.2; .delta.=4.15-3.82, m, 7.times.2H;
.delta.=3.67, t, 1H, J=25.2; .delta.=2.69, t, 2H, J=7.1;
.delta.=1.29 and 1.17, t, 4.times.3H, J=7.1.
[0138] MS (70 eV)
[0139] 552 (M.sup.+, 4), 525 (5), 512 (5), 173 (11), 160 (12), 159
(100), 132 (10), 105 (6), 54 (12).
[0140] IR
[0141] 2983 (m), 2932 (m), 1720 (w), 1600 (m), 1508 (s), 1477 (w),
1391 (w), 1367 (w), 1299 (w), 1250(s), 1164 (w), 1029 (s br),
974(s), 877 (m), 798 (w), 750 (m), 696 (w), 553 (m).
Example 12
Tetraethyl
1-{4-[3-(methyl-pyridin-2-yl-amino)-propoxy]-phenyl}-methyliden-
e-1,1-bisphosphonate
[0142] ##STR18##
[0143] A solution of 2.1 g (4.79 mmol) of tetraethyl
1-[4-(3-hydroxy-propoxy)-phenyl]-methylidene-1,1-bisphosphonate in
20 ml CH.sub.2Cl.sub.2 was added dropwise to a solution of
Dess-Martin reagent (3.1 g, 7.31 mmol) in 20 ml CH.sub.2Cl.sub.2 at
0.degree. C. Stirring was continued at 0.degree. C. for 15 min
followed by warming to room temperature. Reaction progress was
monitoring by GC. After 3 h the mixture was poured into 100 of
saturated bicarbonate solution containing 7.7 g (48.7 mmol) of
Na.sub.2S.sub.2O.sub.3. The aqueous solution was extracted with
CH.sub.2Cl.sub.2 (3.times.30 ml). The organic layers were combined
together, washed with water (3.times.50 ml) and saturated NaCl
solution (2.times.50 ml) and dried over MgSO.sub.4. The crude (1.90
g, 4.35 mmol, yield 90.82%) tetraethyl
1-[4-(3-oxo-propoxy)-phenyl]-methylidene-1,1-bisphosphonate was
concentred by rotary evaporator and used in the next step without
further purification. Acetic acid (2 ml) were added to a solution
of 2.35 g (21.88 mmol) of 2-methylaminopyridine and 1.90 g (4.35
mmol) of tetraethyl
1-[4-(3-oxo-propoxy)-phenyl]-methylidene-1,1-bisphosphonate in 15
ml MeOH. The mixture was stirred for 1 h at room temperature then,
sodium cyanoborohydride was added portionwise (0.55 g, 8.71 mmol).
After addition, stirring was continued overnight. A saturated
bicarbonate solution was added and the mixture taken up into
CH.sub.2Cl.sub.2 (3.times.50 ml) and washed with water (50 ml),
saturated NaCl solution (2.times.50 ml) and dried over MgSO.sub.4.
The oil residue obtained after solvent removal (3.0 g) was purified
by chromatography (Aluminium oxide 90 active neutral,
CH.sub.2Cl.sub.2:MebH 98:2) to give 0.43 g (0.81 mmol,
C.sub.24H.sub.38N.sub.2O.sub.7P.sub.2, M.sub.W=528.53, oil, yield
18.6%) of tetraethyl
1-{4-[3-(methyl-pyridin-2-yl-amino)-propoxy]-phenyl}-methylidene-1,1-bisp-
hosphonate.
[0144] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0145] .delta.=8.14, dxd, 1H, J=5.8, J=2.0; .delta.=7.41, dxdxd,
1H, J=8.6, J=7.0, J=2.0; .delta.=7.38, dxt, 2H, J=8.9, J=2.1;
.delta.=6.86, d, 2H, J=8.7; .delta.=6.53-6.50, m, 2.times.1H;
.delta.=4.16-4.01 and 3.96-3.91, m, 4.times.2H; .delta.=4.00, t,
2H, J=6.1; .delta.=3.73, t, 2H, J=6.9; .delta.=3.67, t, 1H, J=25.2;
.delta.=2.09, quint, 2H, J=6.7; .delta.=1.29 and 1.16, t,
4.times.3H, J=7.1.
[0146] MS (70 eV)
[0147] 528 (M.sup.+, 2), 408 (9), 271 (8), 150 (11), 149 (100), 136
(29), 135 (27), 122 (15), 121 (44), 107 (19), 93 (10), 78 (9), 57
(13).
[0148] IR
[0149] 2982 (m), 2932 (m), 1598 (s), 1560 (w), 1510 (s), 1424 (w),
1389 (w), 1232 (w), 1299 (w), 1251 (s), 1183 (w), 1163 (w), 1028
(s), 973 (s), 876 (m), 711 (w), 552 (m).
Example 13
Tetraisopropyl
1-[4-(pyridin-2-yl-methoxy)-phenyl]-methylidene-1,1-bisphosphonate
[0150] ##STR19##
[0151] Under nitrogen and with exclusion of light, a solution of
0.8 ml (5.07 mmol) of diethyl azodicarboxylate and 0.60 g (5.50
mmol) of 2-hydroxymethyl-pyridine in 30 ml THF was added very
slowly (1 drop/2 sec) to the mixture of 1.28 g (4.88 mmol) of
triphenyl phosphine and 1.74 g (3.99 mmol) of tetraisopropyl
1-(4-hydroxy-phenyl)-methylidene-1,1-bisphosphonate in 40 ml THF at
room temperature. The mixture was stirred at room temperature
overnight. After solvent removal by rotary evaporator, the residue
(5.11 g) was purified by flash chromatography (Silica gel 60,
AcOEt:MeOH 95:5, R.sub.f 0.12) to give 1.16 g (2.20 mmol,
C.sub.25H.sub.39NO.sub.7P.sub.2, M.sub.W=527.54, oil, yield 55.00%)
of tetraisopropyl
1-[4-(pyridin-2-yl-methoxy)-phenyl]-methylidene-1,1-bisphosphonate.
[0152] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0153] .delta.=8.60, d, 1H, J=4.6; .delta.=7.7, t, 1H, J=7.7;
.delta.=7.50, d, 1H, J=7.8; .delta.=7.38, d, 2H, J=8.4;
.delta.=7.23, dxd, 1H, J=7.2, J=5.2; .delta.=6.94, d, 2H, J=8.4;
.delta.=5.20, s, 2H; .delta.=4.74-4.54, m, 4.times.1H;
.delta.=3.56, t, 1H, J=25.3; .delta.=1.29, d, 2.times.3H, J=6.2;
.delta.=1.27-1.23, m, 4.times.3H; .delta.=1.01, d, 2.times.3H,
J=6.2.
[0154] MS (70 eV)
[0155] 528 (4), 527 (M.sup.+, 14), 226 (7), 187 (6), 94 (8), 93
(100), 92 (30), 65 (8).
Example 14
Tetraethyl
2-[4-(pyridin-2-yl-methoxy)-phenyl]-vinylidene-1,1-bisphosphona-
te
[0156] ##STR20##
[0157] Under nitrogen and with exclusion of light, a solution of
3.5 ml (22.17 mmol) of diethyl azodicarboxylate and 2.22 g (20.34
mmol) of 2-hydroxymethyl-pyridine in 10 ml THF was added very
slowly (1 drop/2 sec) to the mixture of 5.62 g (21.43 mmol) of
triphenyl phosphine and 8.0 g (20.39 mmol) of tetraethyl
2-(4-hydroxy-phenyl)-vinylidene-1,1-bisphosphonate in 50 ml THF at
room temperature. The mixture was stirred at room temperature
overnight. After solvent removal by rotary evaporator, the residue
was purified by flash chromatography (Silica gel 60, AcOEt:MeOH
90:10, R.sub.f 0.18) to give 7.43 g (15.37 mmol,
C.sub.22H.sub.31NO.sub.7P.sub.2, M.sub.W=483.44, oil, yield 75.57%)
of tetraethyl
2-[4-(pyridin-2-yl-methoxy)-phenyl]-vinylidene-1,1-bisphosphonate.
[0158] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0159] .delta.=8.61, d, 1H, J=4.9; .delta.=8.22, dxd, 1H, J=48.0,
J=29.4; .delta.=7.84, d, 2H, J=8.8; .delta.=7.72, txd, 1H, J=7.7,
J=1.6; .delta.=7.50, d, 1H, J=7.8; .delta.=7.25, dxd, 1H, J=7.4,
J=5.1; .delta.=7.00, d, 2H, J=8.8; .delta.=5.25, s, 2H;
.delta.=4.22-4.03, m, 4.times.2H; .delta.=1.37 and 1.20,
4.times.3H, J=7.1.
[0160] MS (70 eV)
[0161] 483 (M.sup.+, 10), 391 (20), 374 (12), 347 (22), 346 (100),
293 (9), 199 (16), 93 (71), 92 (46), 65 (18).
Example 15
Tetraethyl 2-(4benzyloxy-phenyl)vinylidene-1,1-bisphosphonate
[0162] ##STR21##
[0163] Titanium(IV) chloride (9.1 g, 47.97 mmol, was added to 40 ml
THF cooled to 0.degree. C. followed by dropwise addition of 3.80 g
(17.90 mmol) of 4-benzyloxybenzaldehyde in 20 ml THF. After
stirring for 45 min at 0.degree. C., a solution of 6.26 g (21.72
mmol) of tetraethyl methylenediphosphonate in 20 ml THF was added
followed, 10 min later, by the addition of 8.69 g (85.9 mmol) of
4-methylmorpholine. The dark resulting mixture was allowed to warm
to room temperature. Water was then added and the solution was
extracted with diethyl ether (3.times.30). The organic layer was
washed with water and saturated NaCl solution and dried over
MgSO.sub.4.
[0164] The final residue (4.38 g) was purified by flash
chromatography (Silica gel 60, AcOEt:MeOH 95:5, R.sub.f 0.27) to
give 2.07 g (4.29 mmol, C.sub.23H.sub.32O.sub.7P.sub.2,
M.sub.W=482.45, oil, yield 23.97%) of tetraethyl
2-(4-benzyloxy-phenyl)-vinylidene-1,1-bisphosphonate.
[0165] .sup.1H-NMR (CDCl.sub.3, 500 MHz) .delta.=8.23, dxd, 1H,
J=48.4, J=29.5; .delta.=7.85, d, 2H, J=8.8; .delta.=7.44-7.32, m,
5H; .delta.=6.99, d, 2H, J=8.8; .delta.=5.12, s, 2H;
.delta.=4.23-4.03, m, 4.times.2H; .delta.=1.38 and 1.21, t,
2.times.3H, J=7.1.
[0166] MS (70 eV)
[0167] 483 (1), 482 (M.sup.+, 6), 346 (10), 345 (45), 92 (8), 91
(100).
Example 16
Diethyl
2-[3-(3-N-phthalimido-propoxy)-phenyl]-vinylidene-1-phosphonate
[0168] ##STR22##
[0169] A solution of 12.12 g (43.85 mmol) of
N-(3-bromopropyl)phthalimide in 50 ml 2-butanone was added dropwise
to a stirred mixture of 5.59 g (43.49 mmol) of
3-hydroxybenzaldehyde, 8.49 g (61.43 mmol) of potassium carbonate
and 1.32 g (3.97 mmol) of tetrabutyl ammonium bromide in 150 ml
2-butanone.
[0170] The reaction mixture was warmed under reflux overnight then
allowed to cool to room temperature. The final mixture was poured
into water and taken up with AcOEt (3.times.50 ml). The organic
layer was washed with saturated NaCl solution, dried over
MgSO.sub.4, filtered and then concentred by rotavapory evaporator.
The residue (17.27 g) was purified by flash chromatography (Silica
gel 60, AcOEt:Hexane 30:70, R.sub.f 0.20) to give 7.29 g (23.57
mmol, C.sub.18H.sub.15NO.sub.4, M.sub.W=309.33, oil, yield 54.20%)
of 3-(3-N-phthalimido-propoxy)-benzaldehyde.
[0171] A solution of 3.5 g (12.14 mmol) of tetraethyl
methylenephosphonate in 30 ml 1,4-dioxane was added to a stirred
suspension of NaH (1.35 g, 33.75 mmol, 60% dispersion in mineral
oil) in 30 ml 1,4-dioxane at 1I0.degree. C. The mixture was allowed
to warm to room temperature and stirred for 30 min then, a solution
of 3.35 g (10.83 mmol) of 3-(3-N-phthalimido-propoxy)-benzaldehyde
in 30 ml 1,4-dioxane was added. The reaction mixture was warmed
under reflux over 4 h.
[0172] The excess of NaH was destroyed by careful treatment with
ethyl acetate and the dioxane was removed by rotary evaporator. A
saturated ammonium chloride solution was then added and the mixture
taken up into AcOEt (4.times.50 ml) and washed with water and
saturated NaCl solution. The organic layer was dried over
MgSO.sub.4 and concentred by rotary evaporator. The residue (3.66
g) was purified by flash chromatography (Silica gel 60, AcOEt,
R.sub.f 0.22) to give 2.48 g (5.59 mmol, C.sub.23H.sub.26NO.sub.6P,
M.sub.W=443.44, oil, yield 54.20%) of
2-[3-(3-N-phthalimido-propoxy)-phenyl]-vinylidene-1-phosphonate.
[0173] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0174] .delta.=7.85-7.84, m, 2H; .delta.=7.74-7.72, m, 2H;
.delta.=7.41, dxd, 1H, J=22.4, J=17.5; .delta.=1.25, t, 1H, J=7.9;
.delta.=7.06, d, 1H, J=7.7; .delta.=6.89, s br, 1H; .delta.=6.82,
dxd, 1H, J=8.1, J=2.3; .delta.=6.18, t, 1H, J=17.5;
.delta.=4.18-4.08, m, 2.times.2H; .delta.=4.05, t, 2H, J=6.8;
.delta.=3.93, t, 2H, J=6.0; .delta.=2.20, quint, 2H, J=6.3;
.delta.=1.36, t, 2.times.3H, J=7.1.
[0175] MS (70 eV)
[0176] 443 (M.sup.+, 21), 283 (15), 269 (40), 189 (49), 188 (100),
161 (20), 160 (72), 147 (11), 133 (12), 130 (42), 118 (11), 105
(10), 104 (13), 102 (10), 91 (11), 77 (19), 76 (12).
Example 17
Diethyl
2-14-3-N-phthalimido-propoxy)-phenyl]-vinylidene-1-phosphonate
[0177] ##STR23##
[0178] A solution of 12.41 g (46.29 mmol) of
N-(3-bromopropyl)phthalimide in 50 ml 2-butanone was added dropwise
to a stirred mixture of 8.99 g (65.05 mmol) of potassium carbonate,
5.59 g (43.49 mmol) of 4-hydroxybenzaldehyde and 1.42 g (4.27 mmol)
of tetrabutyl ammonium bromide in 150 ml 2-butanone.
[0179] The reaction mixture was warmed under reflux over 5.5 h then
allowed to cool to room temperature. The final mixture was poured
into water and taken up with AcOEt (3.times.50 ml). The organic
layer was washed with saturated NaCl solution, dried over
MgSO.sub.4, filtered and then concentred by rotary evaporator.
[0180] After solvent removal, the residue (GC>99.5%) was used in
the next step without fuirther purification. We obtained 12.65 g
(40.90 mmol, C.sub.18H.sub.15NO.sub.4, M.sub.W=309.33, oil, yield
89.36%) of 4-(3-N-phthalimido-propoxy)-benzaldehyde.
[0181] A solution of 7.06 g (24.50 mmol) of tetraethyl
methylenephosphonate in 50 ml 1,4-dioxane was added to a stirred
suspension of NaH (2.80 g, 70.0 mmol, 60% dispersion in mineral
oil) in 50 ml 1,4-dioxane at 10.degree. C. The mixture was allowed
to warm to room temperature and stirred for 30 min then, a solution
of 6.72 g (21.73 mmol) of 4-(3-N-phthalimido-propoxy)-benzaldehyde
in 50 ml 1,4-dioxane was added. The reaction mixture was warmed
under reflux over 2.5 h.
[0182] The excess of NaH was destroyed by careful treatment with
ethyl acetate and the dioxane was removed by rotary evaporator. A
saturated ammonium chloride solution was then added and the mixture
taken up into CH.sub.2Cl.sub.2 (4.times.50 ml) and washed with
water and saturated NaCl solution. The organic layer was dried over
MgSO.sub.4 and concentred by rotary evaporator. The residue (10.54
g) was purified by flash chromatography (Silica gel 60, AcOEt,
R.sub.f 0.18) to give 5.94 g (13.40 mmol,
C.sub.23H.sub.26NO.sub.6P, M.sub.W=443.44, solid, mp 87-89.degree.
C., yield 61.67%) of
2-[3-(3-N-phthalimido-propoxy)-phenyl]-vinylidene-1-phosphonate.
[0183] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0184] .delta.=7.85-7.83, m, 2H; .delta.=7.73-7.71, m, 2H;
.delta.=7.43, dxd, 1H, J=22.6, J=17.5; .delta.=7.40, d, 2H, J=8.6;
.delta.=6.79, d, 2H, J=8.7; .delta.=6.07, t, 1H, J=17.6;
.delta.=4.15-4.08, m, 2.times.2H; .delta.=4.06, t, 2H, J=6.0;
.delta.=3.91, t, 2H, J=6.8; .delta.=2.20, quint, 2H, J=6.3;
.delta.=1.35, t, 2.times.3H, J=7.1.
[0185] MS (70 eV)
[0186] 444 (2), 443 (M.sup.+, 4), 189 (11), 188 (100), 160 (35),
130 (5).
Example 18
Diethyl
2-13-(2-pyridin-2-yl-ethoxy)-phenyl)-vinylidene-1-phosphonate
[0187] ##STR24##
[0188] Under nitrogen and with exclusion of light, a solution of
50.0 ml (0.32 mol) of diethyl azodicarboxylate and 35.7 g (0.29mol)
of 3-hydroxybenzaldehyde in 150 ml THF was added very slowly to the
mixture of 84.0 g (0.32 mol) of triphenyl phosphine and 36.0 g
(0.29 mol) of 2-(2-hydroxyethyl)-pyridine in 400 ml THF at room
temperature. The mixture was stirred at room temperature overnight.
After solvent removal by rotary evaporator, the residue (146.0 g)
was purified by repeated flash chromatography (Silica gel 60,
AcOEt:Hexane 40:60, R.sub.f 0.26) to give 25.2 g (0.11 mol,
C.sub.14H.sub.13NO.sub.2, M.sub.W=227.27, oil, yield 37.93%) of
3-(2-pyridin-2-yl-ethoxy)-benzaldehyde.
[0189] A solution of 5.30 g (18.39 mmol) of tetraethyl
methylenephosphonate in 50 ml 1,4-dioxane was added to a stirred
suspension of NaH (1.49 g, 37.25 mmol, 60% dispersion in mineral
oil) in 50 ml 1,4-dioxane at 10.degree. C. The mixture was allowed
to warm to room temperature and stirred for 30 min then, a solution
of 3.50 g (15.44 mmol) of 3-(2-pyridin-2-yl-ethoxy)-benzaldehyde in
50 ml 1,4-dioxane was added. The reaction mixture was warmed under
reflux over 2.5 h.
[0190] The excess of NaH was destroyed by careful treatment with
ethyl acetate and the dioxane was removed by rotary evaporator. A
saturated ammonium chloride solution was then added and the mixture
taken up into AcOEt (4.times.100 ml) and washed with water and
saturated NaCl solution. The organic layer was dried over
MgSO.sub.4 and concentred by rotary evaporator. The residue (6.59
g) was purified by flash chromatography (Silica gel 60, AcOEt:MeOH
95:5, R.sub.f 0.27) to give 4.03 g (11.15 mmol,
C.sub.19H.sub.24NO.sub.4P, M.sub.W=361.38, oil, yield 72.22%) of
diethyl
2-[3-(2-pyridin-2-yl-ethoxy)-phenyl]-vinylidene-1-phosphonate.
[0191] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0192] .delta.=8.57, d, 1H, J=4.7; .delta.=7.64, txd, 1H, J=7.7,
J=1.8; .delta.=7.45, dxd, J=22.4, J=17.5; .delta.=7.29, d, 1H,
J=7.5; .delta.=7.28, t, 2H, J=7.9; .delta.=7.17, dxd, 1H, J=7.5,
J=4.9; .delta.=7.07, d, 1H, J=7.7; .delta.=7.03, s, 1H;
.delta.=6.93, dxd, 1H, J=8.1, J=2.3; .delta.=6.23, t, 1H, J=17.5;
.delta.=4.39, t, 2H, J=6.6; .delta.=4.16-4.10, m, 2.times.2H;
.delta.=3.28, t, 2H, J=6.6; .delta.=1.35, t, 2.times.3H, J=7.0.
[0193] MS (70 eV)
[0194] 361 (M.sup.+, 6), 269 (9),213 (6), 149 (8),125 (6),122 (10),
107 (10), 106 (100), 93 (43), 78 (8).
Example 19
Tetraethyl
2-[4(pyridin-2-yl-methoxy)phenyl]-ethylidene-1,1-bisphosphonate
[0195] ##STR25##
[0196] Under nitrogen and with exclusion of light, a solution of
1.8 ml (1 1.40 mmol) of diethyl azodicarboxylate and 1.12 g (10.26
mmol) of 2-hydroxymethyl-pyridine in 10 ml THF was added very
slowly (1 drop/2 sec) to the mixture of 2.86 g (10.90 mmol) of
triphenyl phosphine and 4.1 g (10.40 mmol) of tetraethyl
2-(4-hydroxy-phenyl)-ethylidene-1,1-bisphosphonate in 30 ml THF at
room temperature. The mixture was stirred at room temperature
overnight. After solvent removal by rotary evaporator, the residue
was purified by flash chromatography (Silica gel 60, AcOEt:MeOH
95:5, R.sub.f 0.18) to give 1.8 g (3.71 mmol,
C.sub.22H.sub.33NO.sub.7P.sub.2, M.sub.W=485.46, oil, yield 36.16%)
of tetraethyl
2-[4-(pyridin-2-yl-methoxy)-phenyl]-ethylidene-1,1-bisphosphonate.
[0197] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0198] .delta.=8.60, d, 1H, J=4.6; .delta.=7.71, txd, 1H, J=7.7,
J=1.6; .delta.=7.51, d, 1H, J=7.8; .delta.=7.22, dxd, 1H, J=7.2,
J=5.0; .delta.=7.20, d, 2H, J=38.6; .delta.=6.90, d,2H, J=8.6;
.delta.=5.19, s, 2H; .delta.=4.16-4.04, m, 4.times.2H;
.delta.=3.19, txd, 2H, J=16.5, J=6.2; .delta.=2.59, txd, 1H,
J=23.9, J=6.3; .delta.=1.29-1.25, m, 4.times.3H.
[0199] MS (70 eV)
[0200] 485 (M.sup.+, 5), 393 (17), 349 (21), 348 (100), 93 (51), 92
(30), 65 (12).
Example 20
Tetraethyl 1-(4-hydroxy-phenyl)-ethylidene-1,2-bisphosphonate
[0201] ##STR26##
[0202] A solution of 27.12 g (94.09 mmol) of tetraethyl
methylenephosphonate in 100 ml 1,4-dioxane was added to a stirred
suspension of NaH (8.22 g, 205.50 mmol, 60% dispersion in mineral
oil) in 100 ml 1,4-dioxane at 10.degree. C. The mixture was allowed
to warm to room temperature and stirred for 30 min then, a solution
of 17.03 g (80.24 mmol) of 4-benzyloxybenzaldehyde in 100 ml
1,4-dioxane was added. The reaction mixture was warmed under reflux
over 2.5 h. The excess of NaH was destroyed by careful treatment
with ethyl acetate and the dioxane was removed by rotary
evaporator. A saturated ammonium chloride solution was then added
and the mixture taken up into AcOEt (4.times.100 ml) and washed
with water and saturated NaCl solution. The organic layer was dried
over MgSO.sub.4 and concentred by rotary evaporator. The residue
(10.58 g) was purified by flash chromatography (Silica gel 60,
AcOEt, R.sub.f 0.24) to give 5.55 g (16.0 mmol,
C.sub.19H.sub.23O.sub.4P, M.sub.W=346.37, oil, yield 19.94%) of
diethyl 2-(4-benzyloxyphenyl)-vinylidene-1-phosphonate.
[0203] A solution of 7.08 g (51.27 mmol) of diethyl phosphite in 30
ml DME was added to a stirred suspension of NaH (2.78 g, 69.50
mmol, 60% dispersion in mineral oil) in 50 ml DME cooled to
0.degree. C. The mixture was allowed to warm to room temperature
and stirred for 30 min then, a solution of diethyl
2-(4-benzyloxyphenyl)-vinylidene-1-phosphonate (5.55 g, 16.0 mmol)
in 50 ml DME was added. The final mixture was stirred at room
temperature overnight. The excess of NaH was destroyed by careful
treatment with ethyl acetate. Water was then added and the mixture
taken up into CH.sub.2Cl.sub.2 (3.times.100 ml) and washed with
water, saturated NaCl solution and dried over MgSO.sub.4. The
residue (5.69 g) obtained after solvent removal was purified by
flash chromatography (Silica gel 60, AcOEt:MeOH 95:5, R.sub.f 0.21)
to give 1.71 g (3.53 mmol, C.sub.23H.sub.34O.sub.7P.sub.2,
M.sub.W=484.47, oil, yield 22.06%) of tetraethyl
1-(4-benzyloxy-phenyl)-ethylidene-1,2-bisphosphonate.
[0204] 10% Palladium on activated charcoal (1.12 g, 1.05 mmol) was
added to a solution of 1.71 g (3.53 mmol) of tetraethyl
1-(4-benzyloxy-phenyl)-ethylidene-1,2-bisphosphonate in 100 ml
EtOH. The mixture was then submitted to hydrogenation under
pressure (60 p.s.i.) at room temperature for 2 h. After filtration
over MgSO.sub.4, the solution was concentred by rotary evaporator
and the residue (1.52 g) was purified by flash chromatography
(Silica gel 60 , AcOEt:MeOH 90:10, R.sub.f 0.24) to give 1.32 g
(3.35 mmol, C.sub.16H.sub.28O.sub.7P.sub.2, M.sub.W=394.34, oil,
yield 94.90%) of tetraethyl
1-(4-hydroxy-phenyl)-ethylidene-1,2-bisphosphonate.
[0205] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0206] .delta.=9.0-7.3, s br, 1H; .delta.=7.13, dxd, 2H, J=8.6,
J=2.2; .delta.=6.61, d, 2H, J=8.5; .delta.=4.12-3.59, m,
4.times.2H; .delta.=3.43-3.32, m, 1H; .delta.=2.49-2.41, m, 2H;
.delta.=1.31, 1.20, 1.12 and 1.08, t, 4.times.3H, J=7.1.
[0207] MS (70 eV)
[0208] 395 (5), 394 (M.sup.+, 22), 258 (15), 257 (100), 229 (16),
213 (7), 201 (29), 185 (8), 120 (20), 109 (11), 81 (10).
Example 21
Tetraethyl 1-(3-hydroxy-phenyl)ethylidene-1,2-bisphosphonate
[0209] ##STR27##
[0210] A solution of 20.0 g (69.4 mmol) of tetraethyl
methylenephosphonate in 100 ml 1,4-dioxane was added to a stirred
suspension of NaH (8.9 g, 222.5 mmol, 60% dispersion in mineral
oil) in 10 ml 1,4-dioxane at 10.degree. C. The mixture was allowed
to warm to room temperature and stirred for 30 min then, a solution
of 6.0 g (48.15 mmol) of 3-hydroxybenzaldehyde in 100 ml
1,4-dioxane was added. The reaction mixture was warmed under reflux
for 4 h. The excess of NaH was destroyed by careful treatment with
ethyl acetate and the dioxane was removed by rotary evaporator. A
saturated ammonium chloride solution was then added and the mixture
taken up into CH.sub.2Cl.sub.2 (4.times.100 ml) and washed with
water and saturated NaCl solution. The organic layer was dried over
MgSO.sub.4 and concentred by rotary evaporator. The residue (21.05
g) was purified by flash chromatography (Silica gel 60, AcOEt,
R.sub.f 0.20) to give 11.34 g (44.26 mmol,
C.sub.19H.sub.23O.sub.4P, M.sub.W=346.37, oil, yield 91.92%) of
diethyl 2-(3-hydroxyphenyl)-vinylidene-1-phosphonate.
[0211] A solution of 4.5 g (32.59 mmol) of diethyl phosphite in 40
ml DME was added to a stirred suspension of NaH (1.7 g, 42.5 mmol,
60% dispersion in mineral oil) in 40 ml DME cooled to 0.degree. C.
The mixture was allowed to warm to room temperature and stirred for
30 min then, a solution of diethyl
2-(3-hydroxyphenyl)-vinylidene-1-phosphonate (2.46 g, 9.60 mmol) in
40 ml DME was added. The final mixture was stirred at room
temperature overnight. The excess of NaH was destroyed by careful
treatment with ethyl acetate. Water was then added and the mixture
taken up into CH.sub.2Cl.sub.2 (3.times.100 ml) and washed with
water, saturated NaCl solution and dried over MgSO.sub.4. The
residue (5.66 g) obtained after solvent removal was purified by
flash chromatography (Silica gel 60, AcOEt:MeOH 95:5, R.sub.f 0.18)
to give 2.38 g (6.04 mmol, C.sub.16H.sub.28O.sub.7P.sub.2,
M.sub.W=394.34, oil, yield 62.92%) of tetraethyl
1-(3-hydroxy-phenyl)-ethylidene-1,2-bisphosphonate.
[0212] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0213] .delta.=8.58-8.46, s br, 1H; .delta.=7.12, t, 1H, J=7.9;
.delta.=7.00, s, 1H; .delta.=6.84, d, 1H, J=7.5; .delta.=6.72, d,
1H, J=8.1; .delta.=4.13-3.48, m, 4.times.2H; .delta.=3.43-3.33, m,
1H; .delta.=2.47-2.39, m, 2H, .delta.=1.31, t, 3H, J=7.1;
.delta.=1.16, t, 3H, J=7.0; .delta.=1.04, t, 3H, J=7.1;
.delta.=1.00, t, 3H, J=7.1.
[0214] MS (70 eV)
[0215] 395 (6), 394 (M.sup.+, 30), 258 (18), 257 (100), 229 (17),
201 (23), 120 (14), 109 (8), 91 (8), 81 (10).
Example 22
Tetraethyl
1-[3-(3-N-phthalimido-propoxy)-phenyl]-ethylidene1,2-bisphospho-
nate
[0216] ##STR28##
[0217] A solution of 0.98 g (3.66 mmol) of
N-(3-bromopropyl)phthalimide in 20 ml 2-butanone was added dropwise
to a stirred mixture of 0.73 g (5.28 mmol) of potassium carbonate,
1.18 g (3.0 mmol) of tetraethyl
1-(3-hydroxy-phenyl)-ethylidene-1,2-bisphosphonate and 0.15 g (0.47
mmol) of tetrabutyl ammonium bromide in 40 ml 2-butanone. The
reaction mixture was warmed under reflux for 4 h then allowed to
cool to room temperature. The final mixture was poured into water
and taken up with CH.sub.2Cl.sub.2 (3.times.50 ml). The organic
layer was washed with saturated NaCl solution, dried over
MgSO.sub.4, filtered and then concentred by rotavapory evaporator.
The residue (2.02 g) was purified by flash chromatography (Silica
gel 60, AcOEt:MeOH 95:5, R.sub.f 0.20) to give 1.63 g (2.80 mmol,
C.sub.27H.sub.37NO.sub.9P.sub.2, M.sub.W=581.54, oil, yield 93.33%)
of tetraethyl
1-[3-(3-N-phthalimido-propoxy)-phenyl]-ethylidene-1,2-bisphosphonate.
[0218] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0219] .delta.=7.85-7.83, m, 2H; .delta.=7.74-7.72, m, 2H;
.delta.=7.18, t, 1H, J=7.9; .delta.=6.97, d, 1H, J=7.6,
.delta.=6.86, s br, 1H; .delta.=6.70, d, 1H, J=8.2;
.delta.=4.15-3.56, m, 6.times.2H; .delta.=3.44-3.33, m, 1H;
.delta.=2.52-2.36, m, 2H; .delta.=2.18, quint, 2H, J=6.1;
.delta.=1.30, t, 3H, J=7.0; .delta.=1.14, t, 3H, J=7.0;
.delta.=1.10, t, 3H, J=7.0; .delta.=1.05, t, 3H, J=7.0.
[0220] MS (70 eV)
[0221] 582 (6), 581 (M.sup.+, 18), 445 (13), 444 (49), 394 (21),
285 (13), 258 (15), 257 (83), 229 (19), 201 (26), 189 (13), 188
(100), 160 (32), 120 (16), 109 (12), 91 (12), 81 (14).
Example 23
Tetraethyl
1-(4-{2-[(2-cyanoethyl)phenyl-amino]-ethoxy}-phenyl)-ethylidene-
-1,2-bisphosphonate
[0222] ##STR29##
[0223] Under nitrogen and exclusion of light, a solution of 1.0 ml
(6.2 mmol) of diethyl azodicarboxylate and 1.0 g (5.1 mmol) of
N-(2-cyanoethyl)-N-(2-hydroxyethyl)-aniline in 30 ml THF was added
very slowly (1 drop/2 sec) to the mixture of 1.5 g (5.6 mmol) of
triphenyl phosphine and 1.7 g (4.3 mmol) of tetraethyl
1-(4-hydroxy-phenyl)-ethylidene-1,2-bisphosphonate in 30 ml THF at
room temperature. The mixture was stirred at room temperature
overnight. After solvent removal by rotary evaporator, the residue
(5.54 g) was purified by flash chromatography (Aluminium oxide 90
active neutral, CHCl.sub.3:hexane 90:10) to give 0.83 g (1.46 mmol,
C.sub.27H.sub.40N.sub.2O.sub.7P.sub.2, M.sub.W=566.58, oil, yield
33.96%) of tetraethyl
1-(4-{2-[(2-cyano-ethyl)-phenyl-amino]-ethoxy}-phenyl)-ethylidene-1,2-bis-
phosphonate.
[0224] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0225] .delta.=7.31-7.26, m, 4.times.1H; .delta.=6.83, 2H, J=8.6;
.delta.=6.79, t, 1H, J=7.3; .delta.=6.72, d, 2H, J=8.1;
.delta.=4.11, t, 2H, J=5.3; .delta.=4.09-3.57, m, 6.times.2H;
.delta.=3.45-3.33, m, 1H; .delta.=2.68, , t, 2H, J=7.2;
.delta.=2.59-2.34, m, 2H; .delta.=1.30, t, 3H, J=7.1; .delta.=1.15,
t, 3H, J=7.1; .delta.=1.10, t, 3H, J=7.1; .delta.=1.05, t, 3H,
J=7.1.
[0226] MS (70 eV)
[0227] 567 (7), 566 (M.sup.+, 17), 526 (16), 394 (16), 173 (27),
160 (26), 159 (100), 132 (24), 109 (8), 106 (11), 105 (12), 104
(13), 91 (13), 81 (10), 77 (11).
[0228] IR
[0229] 2963 (m), 2932 (m), 2248 (w), 1600 (m), 1508 (m), 1391 (w),
1367 (w), 1245 (s), 1181 (w), 1098 (w), 1029 (s), 969 (m), 794 (m),
750 (m), 696 (w), 542 (w), 507 (m).
Example 24
Tetraethyl
1-[4-(3-N-phthalimidopropoxy)phenyl]-ethylidene-1,2-bisphosphon-
ate
[0230] ##STR30##
[0231] A solution of 1.33 g (4.83 mmol) of
N-(3-bromopropyl)phthalimide in 15 ml 2-butanone was added dropwise
to a stirred mixture of 0.93 g (6.72 mmol) of potassium carbonate,
1.6 g (4.31 mmol) of tetraethyl
1-(4-hydroxy-phenyl)-ethylidene-1,2-bisphosphonate and 0.18 g (0.56
mmol) of tetrabutyl ammonium bromide in 20 ml 2-butanone.
[0232] The reaction mixture was warmed under reflux for 3.5 h then
allowed to cool to room temperature. The final mixture was poured
into water and taken up with CH.sub.2Cl.sub.2 (3.times.100 ml). The
organic layer was washed with saturated NaCl solution, dried over
MgSO.sub.4, filtered and then concentred by rotavapory evaporator.
The residue (2.72 g) was purified by flash chromatography (Silica
gel 60, AcOEt:MeOH 90:10, R.sub.f 0.32) to give 2.2 g (3.78 mmol,
C.sub.27H.sub.37NO.sub.9P.sub.2, M.sub.W=581.54, oil, yield 87.70%)
of tetraethyl
1-[3-(3-N-phthalimido-propoxy)-phenyl]-ethylidene-1,2-bisphosphonate.
[0233] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0234] .delta.=7.85-7.82, m, 2H; .delta.=7.74-7.72, m, 2H;
.delta.=7.26, dxd, 2H, J=8.8, J=2.2; .delta.=6.75, d, 2H, J=8.5;
.delta.=4.10-3.54, m, 6.times.2H; .delta.=3.44-3.34, m, 1H;
.delta.=2.51-2.34, m, 2H; .delta.=2.18, quint, 2H, J=6.3;
.delta.=1.30, t, 3H, J=6.9; .delta.=1.15, t, 3H, J=7.3;
.delta.=1.09, t, 3H, J=7.0; .delta.=1.05, t, 3H, J=6.6.
[0235] MS (70 eV)
[0236] 582 (12), 581 (M.sup.+, 38), 445 (15), 444 (59), 189 (13),
188 (100), 160 (54), 130 (12), 120 (8), 109 (10), 81 (11).
[0237] IR
[0238] 2983 (m), 1773 (w), 1713 (s), 1611 (w), 1513 (m), 1442 (w),
1396 (m), 1372 (w), 1245 (s), 1183 (w), 1029 (s br), 967 (s), 835
(w), 796 (m), 723 (m), 530 (w).
Example 25
Tetraethyl
1-{4-[2-(methyl-pyridin-2-yl-amino)ethoxy]-phenyl}-ethylidene-1-
,2-bisphosphonate
[0239] ##STR31##
[0240] A solution of 7.69 g (50.53 mmol) of
2-(methyl-pyridin-2-yl-amino)-ethanol in 50 ml DMF was added to a
stirred suspension of NaH (2.72 g, 68.0 mmol, 60% dispersion in
mineral oil) in 150 ml DMF cooled to 0.degree. C. The mixture was
allowed to warm to 15.degree. C. and stirred for 1 h then, a
solution of 4-fluorobenzaldehyde (7.14 g, 57.52 mmol) in 50 ml DMF
was added. The final mixture was stirred at room temperature
overnight. The excess of NaH was destroyed by careful treatment
with ethyl acetate. A mixture ice-water (500 g) was then added and
the mixture taken up into AcOEt (4.times.500 ml) and washed with
water, saturated NaCl solution and dried over MgSO.sub.4. The
residue (24.00 g) obtained after solvent removal was purified by
repeated flash chromatography (Silica gel 60, AcOEt:Hexane 40:60,
R.sub.f 0.21) to give 9.89 g (38.59 mmol,
C.sub.15H.sub.16N.sub.2O.sub.2, M.sub.W=256.30, oil, yield 76.37%)
of 4-[2-(methyl-pyridin-2-yl-amino)-ethoxy]-benzaldehyde.
[0241] A solution of 2.85 g (9.89 mmol) of tetraethyl
methylenephosphonate in 30 ml 1,4-dioxane was added to a stirred
suspension of NaH (0.85 g, 21.25 mmol, 60% dispersion in mineral
oil) in 30 ml 1,4-dioxane at 10.degree. C. The mixture was allowed
to warm to room temperature and stirred for 30 min then, a solution
of 2.0 g (7.80 mmol) of
4-[2-(methyl-pyridin-2-yl-amino)-ethoxy]-benzaldehyde in 30 ml
1,4-dioxane was added. The reaction mixture was warmed under reflux
over 1.5 h.
[0242] The excess of NaH was destroyed by careful treatment with
ethyl acetate and the dioxane was removed by rotary evaporator. A
saturated ammonium chloride solution was then added and the mixture
taken up into AcOEt (4.times.50 ml) and washed with water and
saturated NaCl solution. The organic layer was dried over
MgSO.sub.4 and concentred by rotary evaporator. The residue (2.32
g) was purified by flash chromatography (Silica gel 60 Fluka 60752,
AcOEt:MeOH 98:2, R.sub.f 0.27) to give 0.95 g (2.43 mmol,
C.sub.19H.sub.24NO.sub.4P, M.sub.W=361.38, oil, yield 31.15%) of
diethyl
2-{4-[2-(methyl-pyridin-2-yl-amino)-ethoxy]-phenyl}-vinylidene-1-phosphon-
ate.
[0243] A solution of 1.10 g (7.96 mmol) of diethyl phosphite in 10
ml DME was added to a stirred suspension of NaH (0.42 g, 10.5 mmol,
60% dispersion in mineral oil) in 20 ml DME cooled to 0.degree. C.
The mixture was allowed to warm to room temperature and stirred for
30 min then, a solution of diethyl
2-{4-[2-(methyl-pyridin-2-yl-amino)-ethoxy]-phenyl}-vinylidene-1-phosphon-
ate (0.40 g, 1.02 mmol) in 30 ml DME was added. The final mixture
was stirred at room temperature overnight. The excess of NaH was
destroyed by careful treatment with ethyl acetate. Water was then
added and the mixture taken up into CH.sub.2Cl.sub.2 (3.times.50
ml) and washed with water, saturated NaCl solution and dried over
MgSO.sub.4. The residue (1.55 g) obtained after solvent removal was
purified by flash chromatography (Silica gel 60, AcOEt:MeOH 90:10,
R.sub.f 0.24) to give 0.43 g (0.81 mmol,
C.sub.24H.sub.36N.sub.2O.sub.7P.sub.2, M.sub.W=528.53, oil, yield
79.41%) of
1-{4-[2-(methyl-pyridin-2-yl-amino)-ethoxy]-phenyl}-ethylidene-1,2-bispho-
sphonate.
[0244] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0245] .delta.=8.16, dxd, 1H, J=4.7, J=1.9; .delta.=7.46, txd, 1H,
J=7.8, J=1.9; .delta.=7.28, dxd, 2H, J=8.6, J=2.2; .delta.=6.85, d,
2H, J=8.5; .delta.=6.56, dxd, 1H, J=6.7, J=5.1; .delta.=6.55, d,
1H, J=8.6; .delta.=4.16, t, 2H, J=5.8; .delta.=4.10-3.53, m,
4.times.2H; .delta.=3.98, t, 2H, J=5.6; .delta.=3.45-3.33, m, 1H;
.delta.=3.15, s, 1H; .delta.=2.50-2.43, m, 2H; .delta.=1.30, 1.15,
1.09 and 1.04, t, 4.times.3H, J=7.1.
[0246] MS (70 eV)
[0247] 528 (M.sup.+, 11), 422 (8), 421 (22), 136 (21), 135 (27),
135 (41), 122 (17), 121 (100), 108 (14), 81 (18), 79 (15), 78
(35).
Example 26
Diisopropyl
1-(diethoxy-phosphoryl)-1-{4-[2-(methyl-pyridin-2-yl-amino)-ethoxyl-pheny-
l}-ethylidene-2-phosphonate
[0248] ##STR32##
[0249] A solution of 5.8 g (16.7 mmol) of tetraisopropyl
methylenephosphonate in 30 ml 1,4-dioxane was added to a stirred
suspension of NaH (1.4 g, 34.8 mmol, 60% dispersion in mineral oil)
in 50 ml 1,4-dioxane at 10.degree. C. The mixture was allowed to
warm to room temperature and stirred for 30 min then, a solution of
3.3 g (12.9 mmol) of
4-[2-(methyl-pyridin-2-yl-amino)-ethoxy]-benzaldehyde (for the
preparation see synthesis of tetraethyl
1-{4-[2-(methyl-pyridin-2-yl-amino)-ethoxy]-phenyl}-ethylidene-1,2-bispho-
sphonate, in 50 ml 1,4-dioxane was added. The reaction mixture was
warmed under reflux over 2 h. The excess of NaH was destroyed by
careful treatment with ethyl acetate and the dioxane was removed by
rotary evaporator. A saturated ammonium chloride solution was then
added and the mixture taken up into AcOEt (3.times.100 ml) and
washed with water and saturated NaCl solution. The organic layer
was dried over MgSO.sub.4 and concentred by rotary evaporator. The
residue (4.4 g) was purified by flash chromatography (Silica gel
60, AcOEt:MeOH 98:2, R.sub.f 0.34) to give 1.38 g (3.3 mmol,
C.sub.22H.sub.31N.sub.2O.sub.4P, M.sub.W=418.48, oil, yield 25.58%)
of diisopropyl
2-{4-[2-(methyl-pyridin-2-yl-amino)-ethoxy]-phenyl}-vinylidene-1-phosphon-
ate.
[0250] A solution of 3.6 g (25.9 mmol) of diethyl phosphite in 30
ml DME was added to a stirred suspension of NaH (1.3 g, 32.3 mmol,
60% dispersion in mineral oil) in 50 ml DME cooled to 0.degree. C.
The mixture was allowed to warm to room temperature and stirred for
30 min then, a solution of diisopropyl
2-{4-[2-(methyl-pyridin-2-yl-amino)-ethoxy]-phenyl}-vinylidene-1-phosphon-
ate (1.3 g, 3.3 mmol) in 40 ml DME was added. The final mixture was
stirred at room temperature overnight. The excess of NaH was
destroyed by careful treatment with ethyl acetate. Water was then
added and the mixture taken up into CH.sub.2Cl.sub.2 (3.times.50
ml) and washed with water, saturated NaCl solution and dried over
MgSO.sub.4. The residue (1.8 g) obtained after solvent removal was
purified by flash chromatography (Silica gel 60, AcOEt:MeOH 90:10,
R.sub.f 0.26) to give 0.8 g (1.44 mmol,
C.sub.26H.sub.42N.sub.2O.sub.7P.sub.2, M.sub.W=556.57, oil, yield
43.64%) of diisopropyl
1-(diethoxy-phosphoryl)-1-{4-[2-(methyl-pyridin-2-yl-amino)-ethoxy]-pheny-
l}-ethylidene-2-phosphonate.
[0251] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0252] .delta.=8.15, dxd, 1H, J=5.0, J=1.9; .delta.=7.47, dxdxd,
1H, J=8.5, J=6.9, J=1.9; .delta.=7.29-7.27, m, 2H;
.delta.=6.86-6.84, m, 2H; .delta.=6.53, dxd, 1H, J=7.0, J=5.0;
.delta.=6.53, d, 1H, J=8.6; .delta.=4.16, t, 2H, J=5.8;
.delta.=4.15-3.99 and 3.94-3.30, m, 2.times.2H and 3.times.1H;
.delta.=3.97, t, 2H, J=5.8; .delta.=3.14, s, 3H; .delta.=2.51-2.33,
m, 2H; .delta.=1.35-0.98, m, 6.times.3H.
[0253] MS (70 eV)
[0254] 556 (M.sup.+, <1), 179 (5), 136 (10), 135 (100), 122 (6),
121 (52), 119 (4), 104 (4), 78 (5).
[0255] IR
[0256] 2981 (m), 2932 (m), 1736 (w), 1654 (w), 1598 (m), 1560 (w),
1511 (s), 1426 (w), 1387 (w), 1324 (w), 1245 (s), 1180 (w), 1162
(w), 1099 (w), 1030 (s br), 853 (w), 772 (m), 507 (m).
Example 27
Tetraethyl
1-[3-(2-pyridin-2-yl-ethoxy)-phenyl]-ethylidene-1,2-bisphosphon-
ate
[0257] ##STR33##
[0258] A solution of 2.42 g (17.52 mmol) of diethyl phosphite in 10
ml DME was added to a stirred suspension of NaH (0.88 g, 22.00
mmol, 60% dispersion in mineral oil) in 20 ml DME cooled to
0.degree. C. The mixture was allowed to warm to room temperature
and stirred for 30 min then, a solution of diethyl
2-[3-(2-pyridin-2-yl-ethoxy)-phenyl]-vinylidene-1-phosphonate (2.00
g, 5.53 mmol) in 20 ml DME was added. The final mixture was stirred
at room temperature overnight. The excess of NaH was destroyed by
careful treatment with ethyl acetate. Water was then added and the
mixture taken up into CH.sub.2Cl.sub.2 (3.times.50 ml) and washed
with water, saturated NaCl solution and dried over MgSO.sub.4. The
residue (2.74 g) obtained after solvent removal was purified by
flash chromatography (Silica gel 60, AcOEt:MeOH 98:2, R.sub.f 0.34)
to give 0.47 g (0.94 mmol, C.sub.23H.sub.35NO.sub.7P.sub.2,
M.sub.W=499.49, oil, yield 17.0%) of tetraethyl
1-[3-(2-pyridin-2-yl-ethoxy)-phenyl]-ethylidene-1,2-bisphosphonate.
[0259] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0260] .delta.=8.56, d, 1H, J=4.8; .delta.=7.64, txd, 1H, J=7.6,
J=1.8; .delta.=7.28, d, 1H, J=8.0; .delta.=7.20, t, 1H, J=7.9;
.delta.=7.16, dxd, 1H, J=7.5, J=4.9; .delta.=6.97, d, 1H, J=7.6;
.delta.=6.95, s, 1H; .delta.=6.81, d, 1H, J=8.2; .delta.=4.35, t,
2H, J=6.7; .delta.=4.12-3.54, m, 4.times.2H; .delta.=3.46-3.35, m,
1H; .delta.=3.26, t, 2H, J=6.7; .delta.=2.52-2.38, m, 2H;
.delta.=1.30, 1.12, 1.08 and 1.02, 4.times.3H, J=7.1.
[0261] MS (70 eV)
[0262] 499 (M.sup.+, 6), 408 (14), 407 (77), 363 (11), 362 (50),
315 (17), 257 (12), 165 (14), 109 (11), 107 (15), 106 (100), 93
(25), 81 (11), 71 (12), 57 (21).
Example 28
Tetraethyl
1-(3,5-di-tert-butyl-4-hydroxy-phenyl)-ethylidene-1,2-bisphosph-
onate
[0263] ##STR34##
[0264] Triethylphosphite (29.9 g, 180.0 mmol) was added to 15.3 g
(60.0 mmol) of 2,6-di-tert-butyl-4-chloromethyl-phenol. The mixture
was heated at 120.degree. C. for 3 h then, the excess of reagent
and side products were removed under vacuum. The crude diethyl
1-(3,5-di-tert-butyl-4-hydroxy-phenyl)-methylidene-1-phosphonate
(22.8 g, yield >99%) was used without further purification
(GC>96%). Trifluoromethanesulfonyl chloride was added to a
suspension of NaH (2.1 g, 51.2 mmol, 60% dispersion in mineral oil)
in 75 ml Et.sub.2O at -40.degree. C. (CH.sub.3CN/CO.sub.2). A
solution of 6.62 g (39.4 mmol) of diethyl hydroxymethylphosphonate
in 15 ml Et.sub.2O was added dropwise at the previous mixture kept
at <-30.degree. C. The resulting mixture was stirred at
-40.degree. C. for 2 h, then poured into a saturated NaHCO.sub.3
solution and extracted with CH.sub.2Cl.sub.2 and washed with water,
saturated NaCl solution and dried over MgSO.sub.4. The crude
diethyl phosphonomethyltriflate (12.93 g, yield >99%) obtained
after solvent removal was used without further purification
(GC>99%).
[0265] A volume of 35 ml of a 1.6M solution of n-butyl lithium in
hexane (55.5 mmol) were added to 75 ml THF kept at -78.degree. C.
Diisopropyl amine (5.6 g, 55.5 mmol) was added, the mixture was
stirred for 15 min at -78.degree. C., then a solution of 7.9 g
(22.2 mmol) of diethyl
1-(3,5-di-tert-butyl-4-hydroxy-phenyl)-methylidene-1-phosphonate in
25 ml THF was added dropwise. After a further 15 min stirring at
-78.degree. C. diethyl phosphonomethyltriflate (7.0 g, 23.3 mmol)
was added and the resulting mixture was stirred at -78.degree. C.
for 1h. The cooling bath was removed, the mixture allowed to warm
at room temperature. 100 ml of HCl 10% were then added and the
mixture taken up into CH.sub.2Cl.sub.2 and washed with water,
saturated NaCl solution and dried over MgSO.sub.4. The residue
(17.8 g) obtained after solvent removal was purified by flash
chromatography (Silica gel 60, AcOEt:MeOH 50:50, R.sub.f 0.58) to
give 8.2 g (16.2 mmol, C.sub.24H.sub.44O.sub.7P.sub.2,
M.sub.W=506.56, oil, yield 72.97%) of tetraethyl
1-(3,5-di-tert-butylhydroxy-phenyl)-ethylidene-1,2-bisphosphonate.
[0266] .sup.1H-NMR (CDCl.sub.3, 500 MHz)
[0267] .delta.=7.16, d, 2H, J=2.2; .delta.=5.15, s, 1H;
.delta.=4.13-3.47, m, 4.times.2H; .delta.=3.41-3.31, m, 1H;
.delta.=2.47-2.36, m, 2H; .delta.=1.42, s, 6.times.3H;
.delta.=1.30, t, 3H, J=7.1; .delta.=1.10, t, 3H, J=7.1;
.delta.=1.04, t, 3H, J=7.0; .delta.=1.00, t, 3H, J=7.1.
[0268] MS (70 eV)
[0269] 507 (31), 506 (M.sup.+, 100), 505 (8), 492 (6), 491 (21),
465 (6), 464 (23), 426 (13), 370 (22), 369 (100), 368 (8), 313
(20), 269 (11), 266 (10), 241 (8), 232 (6), 201 (9), 109 (8), 57
(32).
Example 29
Further methylidene1,1-bisphosphonates of Formula (Ia):
[0270] ##STR35## ##STR36##
Example 30
Further alkylidene-1,1-bisphosphonates of Formula (Ib):
[0271] ##STR37##
Example 31
Further alkenylidene-1,1-bisphosphonates of Formula (Ic):
[0272] ##STR38##
Example 32
Further ethylidene-1,2-bisphosphonates of Formula (Ie):
[0273] ##STR39##
Example 33
Biological Activity
[0274] A. Methods
[0275] THP-1 cell line (ATCC TIB-202) are cultured in RPMI 1640
(with 2 mM of L-glutamine and 2 g/l of glucose, Invitrogen)
supplemented with 2 g/l of sodium hydrogen carbonate (Fluka), 10
U/ml of penicillin, 10 .mu.g/ml of streptomycin
(Penicillin-Streptomycin, Invitrogen), 20 .mu.M of
2-mercaptoethanol (Fluka) and 10% FCS (Amimed). Cells are incubated
at 37.degree. C. and 5% CO.sub.2.
[0276] Cells are seeded in 24 well tissue culture plates (Falcon),
2.times.10.sup.5 cells in 500 .mu.l of culture medium per well or 6
well plates for RT-PCR purpose, 1.times.10.sup.6 cells in 2.5 ml of
medium. PMA (Phorbol 12-myristate 13-acetate, Alexis), (stock in
DMSO (Fluka) at 1 mg/ml) and other compounds are diluted in ethanol
and added to the cells at an ethanol final concentration not
exceeding 1%. Same volume of ethanol is added in controls. Plates
are incubated for 3 days at 37.degree. C. and 5% CO.sub.2 to allow
cell differentiation and apoE secretion.
[0277] Medium contained in the wells is harvested and centrifuged 5
min at 1200 rpm. Supernatants are store at -20.degree. C. until
apoE quantification. The cell pellets are resuspended in PBS and
counted with a ZI Coulter Counter. After a wash with PBS, adherent
cells in the well bottoms are detached with trypsin-EDTA solution
10.times. (Invitrogen) diluted in PBS. The reaction is stopped with
the addition of culture medium and the detached cells are
counted.
[0278] ApoE was quantified by ELISA. 96 well plates (Costar) are
coated with 5% of gelatine (from porcine skin, Fluka) in
carbonate-bicarbonate buffer (Sigma) for 1 hour at 37.degree. C.
After removing the coating solution, 100 .mu.l of THP-1 supernatant
are added per well, diluted 5 times in buffer (PBS, 1% Top-Block
(Juro), 0.1% Tween 20 (Fluka)). The incubation last 1 hour at
37.degree. C., then the wells are washed 3 times with 200 .mu.l of
buffer. Plates are incubated for 1 hour at 37.degree. C. with
continuous stirring with the primary antibody (goat anti-human apoE
IgG, Calbiochem) at a 10000-time dilution in buffer, 100 .mu.l per
well. After 3 washes, plates are incubated with the secondary
antibody (rabbit anti-goat-IgG peroxidase conjugated, Sigma)
diluted 5000 times, 100 .mu.l per well, at 37.degree. C. with
shaking. Then the wells are washed 5 more times and the detection
is achieved by adding 100 .mu.l per well of o-phenylenediamine
dihydrochloride (Sigma) and incubated for 15-20 minutes with
shaking at room temperature. When the appropriated colour is
reached, the reaction is stopped by adding 50 .mu.l per well of 3 M
sulfuric acid (Fluka) with shaking for 1 minute at room
temperature. The absorbance is read at 492 nm versus 620 nm with a
microplate photometer (Anthos Reader 2001). TABLE-US-00001 TABLE 1
1,1-Bisphosphonates % ApoE Change 1.25 nM 2.5 nM Compound .mu.M PMA
PMA Tetraisopropyl 1-(4-hydroxy-phenyl)- 25 290 52
methylidene-1,1-bisphosphonate 50 399 35 100 488 73 Tetraethyl
1-(4-hydroxy-phenyl)- 25 -13 -3 methylidene-1,1-bisphosphonate 50
26 -9 100 125 60 Tetramethyl 1-(4-hydroxy-phenyl)- 25 -7 -17
methylidene-1,1-bisphosphonate 50 -27 -25 100 -19 -10 Tetraethyl
1-[4-(3-N-phthalimido- 25 2134 352
propoxy)-phenyl]-methylidene-1,1- 50 2728 405 bisphosphonate 100
3197 482 Tetraethyl 1-{4-[3-(methyl- 25 375 557
pyridin-2-yl-amino)-propoxy]- 50 1197 1165 phenyl}-methylidene-1,1-
100 2431 1696 bisphosphonate Tetraethyl 1-(4-{2-[(2-cyano-ethyl)-
25 276 209 phenyl-amino]-ethoxy}-phenyl)- 50 678 311
methylidene-1,1-bisphosphonate 100 1284 499 Tetraethyl
1-[4-(4-cyano-benzyloxy)- 25 91 134 phenyl]-methylidene-1,1- 50 192
225 bisphosphonate 100 447 328 Tetraethyl
1-[4-(pyridin-2-yl-methoxy)- 25 16 96
phenyl]-methylidene-1,1-bisphosphonate 50 146 192 100 443 574
Tetraethyl 1-[4-(3-pyridin-3-yl-propoxy)- 25 63 136
phenyl]-methylidene-1,1-bisphosphonate 50 103 188 100 337 331
Tetraethyl 1-[4-(2-pyridin-2-yl-ethoxy)- 25 57 38
phenyl]-methylidene-1,1-bisphosphonate 50 133 73 100 363 145
Tetraethyl 1-[4-(2-N-succinimido-ethoxy)- 25 -51 30
phenyl]-methylidene-1,1-bisphosphonate 50 53 70 100 123 132
Tetraethyl 1-[4-(trans, trans-3,7,11- 2.5 14 27
trimethyl-dodeca-2,6,10-trienyloxy)- 5.0 46 47
phenyl]-methylidene-1,1-bisphosphonate 10 31 40 Tetraethyl
1-[4-(2-N-pyrrolidino-ethoxy)- 25 -5 2
phenyl]-methylidene-1,1-bisphosphonate 50 2 7 100 56 32 Tetraethyl
1-[4-(2-N-morpholino-ethoxy)- 25 -17 -20
phenyl]-methylidene-1,1-bisphosphonate 50 -8 -27 100 43 -3
Tetraethyl 1-[4-(2-N-piperidino-ethoxy)- 25 -22 -2
phenyl]-methylidene-1,1-bisphosphonate 50 6 0 100 65 33 Tetraethyl
1-[4-(3-hydroxy-propoxy)- 25 -27 -15
phenyl]-methylidene-1,1-bisphosphonate 50 -21 -16 100 20 0
[0279] TABLE-US-00002 TABLE 2 1,2-Bisphosphonates % ApoE Change
1.25 nM 2.5 nM Compound .mu.M PMA PMA Tetraethyl
1-[4-(3-N-phthalimido- 25 1090 644 propoxy)-phenyl]-ethylidene-1,2-
50 1756 942 bisphosphonate 100 2705 1096 Tetraethyl
1-{4-[2-(methyl-pyridin- 25 77 110
2-yl-amino)-ethoxy]-phenyl}-ethylidene- 50 202 124
1,2-bisphosphonate 100 569 206 Tetraethyl
1-[3-(2-pyridin-2-yl-ethoxy)- 25 31 110
phenyl]-ethylidene-1,2-bisphosphonate 50 51 124 100 274 206
Tetraethyl 1-[4-(pyridin-2-yl-methoxy)- 25 -- 152
phenyl]-ethylidene-1,2-bisphosphonate 50 -- 158 100 -- 289
Tetraethyl 1-(4-ethoxyphenyl)-ethylidene- 25 -16 5
1,2-bisphosphonate 50 -12 4 100 36 54
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